CN112209980B

CN112209980B - substituted purine nucleotides

Info

Publication number: CN112209980B
Application number: CN202010812826.3A
Authority: CN
Inventors: J-P·索玛迪西; A·莫萨
Original assignee: Atea Pharmaceuticals Inc
Current assignee: Atea Pharmaceuticals Inc
Priority date: 2015-03-06
Filing date: 2016-03-07
Publication date: 2023-12-05
Anticipated expiration: 2036-03-07
Also published as: MX2022015612A; CA3182565A1; MY190867A; MX2017011386A; JP7053754B2; US10875885B2; CN107427530A; AU2018282469B2; US20220267366A1; JP2021008494A; US10005811B2; NZ734908A; SA517382236B1; GEP20247600B; HK1247829A1; PH12017501598A1; AU2016229966A1; US20160257706A1; IL254111B; US20200331956A1

Abstract

The present application relates to substituted purine nucleotides. In particular, the present application relates to compounds of structure (a) or pharmaceutically acceptable salts or compositions thereof, for use in treating a host infected with or exposed to HCV virus or other conditions described more fully herein.

Description

Substituted purine nucleotides

The present application is a divisional application of patent application of the application number 201680021594.1, entitled "beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N6-substituted purine nucleotide for HCV treatment", which is 3/7 of the application date.

Priority

The application claims priority from U.S. S. N.62/129,319 submitted on month 5, 3, 6, U.S. S. N.62/253,958 submitted on month 11, 2015, and U.S. S.N 62/276,597 submitted on month 8, 2016, each of which is incorporated herein in its entirety.

Technical Field

The present application relates to nucleotide compounds and compositions, and their use for the treatment of hepatitis c virus ("HCV").

Background

Hepatitis C (HCV) is an RNA single stranded virus and is a member of the genus hepatitis c virus. It is estimated that 75% of all liver disease cases are caused by HCV. HCV infection can lead to cirrhosis and liver cancer, and if kept developing, can lead to liver failure requiring liver transplantation. About 1.7-2 million people worldwide are infected, with an estimated 3-4 million people in America.

RNA polymerase is a key component in targeting RNA single stranded viruses. The HCV nonstructural protein NS5B RNA-dependent RNA polymerase is a key enzyme responsible for initiating and catalyzing viral RNA synthesis. Thus, HCV NS5B is an attractive target for drug research and development of current anti-HCV agents. There are two major subclasses of NS5B inhibitors: nucleoside analogs, which are anabolized to their active triphosphates (which act as alternative substrates for the polymerase); and non-nucleoside inhibitors (NNIs) that bind to the allosteric region of the protein. The nucleoside or nucleotide inhibitor mimics the natural polymerase substrate and acts as a chain terminator. They inhibit the initiation of RNA transcription and the elongation of primary DNA strands.

In addition to targeting RNA polymerase, other RNA viral proteins may also be targeted in combination therapy. For example, another target HCV protein for therapeutic methods is NS3/4A (a serine protease) and NS5A (a non-structural protein that is an essential component of HCV replicase and plays a range of roles for the cellular pathway). .

At month 12 2013, the first nucleoside NS5B polymerase inhibitor sofosbuvir was approvedGilead Sciences)。Is a uracil nucleoside phosphoramidate prodrug, which is taken up by liver cells and activated in the cells to obtain an active metabolite; 2' -deoxy-2 ' - α -fluoro- β -C-methyluridine-5 ' -triphosphate; see the following structure:

Is the first drug that has proven safe and effective in treating some types of HCV infection without the need for co-administration of interferon.Is the third drug to obtain FDA approved breakthrough therapy approval (breakthrough therapy designation).

In 2014, the U.S. FDA approved(ledispasvir, an NS5A inhibitor, and sofosbuvir) for the treatment of chronic hepatitis c virus genotype 1 infection.Is the first combination pill approved for the treatment of chronic HCV genotype 1 infection. It is also the first approved regimen that does not require administration of interferon or ribavirin. In addition, the FDA approved cimaprevir (Olysio) ^TM ) And Sofossa Wei Zuge->As a once daily, total oral, interferon-free and ribavirin treatment regimen for genotype 1HCV infected adults.

In 2014, abbVie's VIEKIRA Pak was also approved by the United states FDA ^TM A multi-pill package comprising dacarbavir (a non-nucleoside NS5B polymerase inhibitor), ombitasvir (an NS5A inhibitor), paritaprevir (an NS3/4A inhibitor) and ritonavir. VIEKIRA Pak ^TM Can be used with or without ribavirin to treat genotype 1 HCV-infected patients, including patients with compensated liver cirrhosis. VIEKIRA Pak ^TM No interferon combination therapy is required.

7 months 2015, U.S. FDA approved Technivie ^TM And Dakliza ^TM For the treatment of HCV genotype 4 and HCV genotype 3, respectively. Technivie ^TM (Ombitasvir/paritaprevir/ritonavir) is approved in combination with ribavirin for treatment of HCV genotype 4 in patients without scarring and cirrhosis and is the first option for HCV-4 infected patients who do not require co-administration with interferon. Dakliza ^TM Is approved toTogether for the treatment of HCV genotype 3 infection. Dakliza ^TM Is the first drug that has proven safe and effective in treating HCV genotype 3 without the need for co-administration of interferon or ribavirin.

10 months 2015, the U.S. FDA warns HCV treatment of Viekira Pak and Technivie may cause severe liver damage, mainly in patients with potential advanced liver disease, and requires additional information to be added to the specification regarding safety. .

Other currently approved therapeutic agents for HCV include interferon alpha-2 b or pegylated interferon alpha-2 bIt can be combined with ribavirin +.>NS3/4A telaprevir (+)>Vertex and Johnson&Johnson), boceprevir (Victrelis) ^TM Merck), cimaprevir (Olysio) ^TM ,Johnson&Johnson), paritaprevir (AbbVie), ombitasvir (AbbVie), (NNI) Dakatavir (ABT-333) and Zepatier from merck ^TM (single tablet combination of two drugs, grazoprevir and elbasvir).

Another NS5B polymerase inhibitor is currently under development. The merck company is developing the uridine nucleotide prodrug MK-3682 (formerly Idenix IDX 21437). The drug is currently in phase II combination testing.

U.S. patents and WO applications describing nucleoside polymerase inhibitors for the treatment of flaviviridae (including HCV) include those filed by the following companies: idenix Pharmaceuticals (6,812,219; 6,914,054;7,105,493;7,138,376;7,148,206;7,157,441;7,163,929;7,169,766;7,192,936;7,365,057;7,384,924;7,456,155;7,547,704;7,582,618;7,608,597;7,608,600;7,625,875;7,635,689;7,662,798;7,824,851;7,902,202;7,932,240;7,951,789;8,193,372;8,299,038;8,343,937;8,362,068;8,507,460;8,637,475,085; 8,680,071;8,691,8,8,8,742,101,951,785; 7,985; 7,001; 2015,025; 2015,2015,2015; 2015,2015,2015,2015,2012/wo/2015,2015,2015; 2015,2012/wo/2015,2015,2015,2015,2015; 2015,2015,2015,2015/wo/2015,2015; merck (6,777,395; 7,105,499;7,125,855;7,202,224;7,323,449;7,339,054;7,534,767;7,632,821;7,879,815;8,071,568;8,148,349;8,470,834;8,481,712;8,541,434;8,697,694;8,715,638,9,061,041;9,156,872 and WO/2013/009737); university of Emerri (Emory) (6,348,587;6,911,424;7,307,065;7,495,006;7,662,938;7,772,208;8,114,994;8,168,583;8,609,627;US 2014/0212382; and WO 2014/1244430); jileydig science/drug company (Gilead Sciences/Pharmasset Inc.) (7,842,672; 7,973,013;8,008,264;8,012,941;8,012,942;8,318,682;8,324,179;8,415,308;8,455,451;8,563,530;8,841,275;8,853,171;8,871,785;8,877,733;8,889,159;8,906,880;8,912,321;8,957,045;8,957; 957,046;9,045,520;9,085,573;9,090,642; and 9,139,604) and (6,908,924; 6,949,522;7,094,094; 7,211,570;7,572; 7,601,820;7,638,638,790; 7,790; 7,790,790; 171,171,208; 8,263,263,263,263, 3,263,735; 8,213,213,642; 8,642,642,75; and (3,735) respectively; 9,642,213,642,924); roche (Hoffman La-Roche) (6,660,721), roche (Roche) (6,784,166; 7,608,599,7,608,601 and 8,071,567); arios biopharmaceutical company (Alios BioPharma Inc) (8,895,723;8,877,731;8,871,737,8,846,896,8,772,474;8,980,865;9,012,427;US 2015/0105341; US 2015/0011497; US 2010/0249068; US 2012/007011; WO 2015/054465; WO 2014/20979; WO 2014/100505; WO 2014/100498; WO 2013/142159; WO 2013/142157; WO 2013/096680; WO 2013/088155; WO 2010/108135), enanta Pharmaceuticals (US 8,575,119;8,846,638;9,085,599;WO 2013/044030; WO 2012/125900), biota (7, 268,119;7,285,658;7,713,941;8,119,607;8,415,309;8,501,699 and 8,802,840), biological crystal company (Biocryst Pharmaceuticals) (7, 388,002;7,429,571;7,514,410;7,560,434;7,994,139;8,133,870;8,163,703, 8,242,085 and 8,440,813), alla 393, LLC (8,889,701 and WO 2015/053662), inhibit ex (8,759,318 and WO/2012/092484), yansen (Janssen Products) (8, 399,429;8,431,588,8,481,510,8,552,021,8,933,052;9,006,29 and 9,012,428), the University of Georgia Foundation (6, 348,587;7,307,065;7,662,938;8,168,583;8,673,926,8,816,074;8,921,384 and 8,946,244), RFS Pharma, LLC (8,895,531;8,859,595;8,815,829;8,609,627;7,560,550;US 2014/0066395; US 2014/02355566; US 2010/0279969; WO/2010/096 and WO/158811), university College Cardiff Consultants Limited (WO/201424/6490; WO 2010/081082; WO/2008/138206), inc (WO/2014/9278 and WO 2014/1680), inc (In/9278 and WO 2014/1680), ind (45/913), pharma, and RFS Pharma (45/3535) Catabasis (WO 2013/090420) and the university of Minnesota board of board (the Regents of the University of Minnesota) (WO 2006/004637).

Nevertheless, there remains a strong medical need to develop safe, effective and well-tolerated anti-HCV therapeutics. The important requirement is expected to be drug resistance. For patients infected with all HCV genotypes, a more effective direct acting antiviral drug can significantly shorten treatment duration and increase compliance and SVR rate. .

Accordingly, it is an object of the present invention to provide compounds, pharmaceutical compositions, and methods and uses for the treatment and/or prevention of HCV infection.

Brief description of the invention

It has been found that formula I, formula II, formula III, formula IV, formula V, formula VICompounds of formula VII, including beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-N ⁶ - (mono-or di-methyl) purine nucleotides, when administered in effective amounts to a host in need thereof, are highly effective against HCV virus. The host may be a human or any animal carrying the virus infection.

The disclosed nucleotides include those having nanomolar activity against HCV in vitro and therapeutic indices ranging up to 25,000 or more.

Surprisingly, prior to the present invention, parent N of the disclosed compounds ⁶ - (methyl) purine nucleosides have not been developed or specifically disclosed as drug candidates. For example, 3' -azido-N was reported in 2010 ⁶ Dimethyl-2, 6-diaminopurine cannot be substantially deaminated by adenosine deaminase over a long period of time (120 minutes), for which reason it is not considered a suitable compound for derivatization as a drug (see e.g. WO 2010/091386, page 86 and corresponding us patent 8,609,627).

However, it has now been found that the compounds of the invention are anabolized to N ⁶ 5-monophosphate esters of substituted purines with substantially no N ⁶ Deamination, which is then anabolized at the 6-position, produces active guanine triphosphate compounds in a manner that provides superior activity and therapeutic index.

In particular, it has been found that β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ -methyl-2, 6-diaminopurine nucleotides and beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -dimethyl-2, 6-diaminopurine nucleotides and other beta-D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N as disclosed below ⁶ 5' -stable phosphate prodrugs or derivatives of substituted purine nucleotides are highly potent against HCV. This is particularly surprising because the parent nucleoside β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ Methyl-2, 6-diaminopurine in replicon assays (EC ₅₀ =15.7 micromolar) indicates that it is unsuitable as a human medicament because of insufficient activity (in combination with reference WO 2010/091386, page 86 and corresponding us patent 8,609,627, indicating N ⁶ -methyl-2, 6-diaminopurineDoes not deaminate in vivo), however the stable racemic phosphate prodrug (phosphoramidate) shows EC in replicon assays ₅₀ =26 nanomole (nM), its activity is increased by at least 600-fold. The corresponding (S) -phosphoramidates exhibit EC ₅₀ =4 nM, an increase in activity of at least 3,900 fold; see structures below and compounds 5-2 in table 7. Due to TC ₅₀ Greater than one hundred micromoles, the compound therefore has a therapeutic index greater than 25,000. In contrast, sofosbuvir has EC ₅₀ ＝53nM，TC ₅₀ Greater than one hundred micromoles and a therapeutic index greater than 1,920..

Likewise, the parent nucleoside β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ -dimethyl-2, 6-diaminopurine in replicon assays (EC ₅₀ Activity in =10.7 micromolar, "μΜ") suggests that it is also unsuitable as a human drug due to insufficient activity, however the stable racemic phosphate prodrug (phosphoramidate) shows EC in replicon assays ₅₀ =12 nM, with an increase in activity of more than 890 fold. The corresponding (S) -phosphoramidate (Compound 25, table 7) also shows EC ₅₀ =4 nM, an increase in activity of at least 2,600 fold; see structure below. In addition, compound 25 also has a therapeutic index greater than 25,000.

In another example, in the replicon assay, the compound ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-N-cyclopropyl-amino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester showed EC ₅₀ =7 nM, the corresponding (S) -phosphoramidate shows EC ₅₀ =5 nM; see compound 27 in table 7 and the structures below. .

As described above, the phosphoramidate is present as beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ Metabolism of methyl-2, 6-diaminopurine nucleosides involves the production of 5' -monophosphates, followed by N ⁶ -methyl-2, 6-diaminopurine base anabolism to produce β -D-2 '-deoxy-2' - α -fluoro-2 '- β -methylguanosine as 5' -monophosphate. The monophosphate is then further anabolized to an active species; 5' -triphosphates. beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-guanine triphosphate with IC against HCV genotype 1b NS5B polymerase ₅₀ ＝0.15μM。

Thus, in one embodiment, the invention is:

wherein the ratio is

Y is NR ¹ R ² ；

R ¹ Is C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CH ₂ F、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ 、CF ₂ CH ₃ And CF (compact F) ₂ CF ₃ )、C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl), -OR ²⁵ 、-C(O)R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ Each of which may be optionally substituted;

R ² is hydrogen, C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ )、–(C ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、–(C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (heteroaryl); and is also provided with

Wherein R is ¹ And R is ² At least one of them is methyl, CH ₂ F、CHF ₂ Or CF (CF) ₃ ；

R ³ Is hydrogen,Bisphosphates, triphosphates, optionally substituted carbonyl-linked amino acids or-C (O) R ^3C ；

R ^3A May be selected from-O-, OH, -O-optionally substituted aryl, -O-optionally substituted heteroaryl or optionally substituted heterocyclyl;

R ^3B may be selected from O-, OH-, optionally substituted N-linked amino acids or optionally substituted N-linked amino acid esters;

R ^3C is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) or-O- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

R ⁴ is a monophosphate, diphosphate, triphosphate, or stable phosphate prodrug, including but not limited to phosphoramidate, phosphorothioate, or any other moiety that is metabolized to a monophosphate, diphosphate, or triphosphate in the host human or animal body; or (b)

R ³ And R is ⁴ Together with the oxygen to which they bind, may form 3',5' -cyclic prodrugs, including but not limited to 3',5' -cyclic phosphate prodrugs;

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl.

In one embodiment, the invention is

Wherein:

y is NR ¹ R ² ；

R ² is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the And

R ^3A Can be selected from O ^- OH, -O-optionally substituted aryl, -O-optionally substituted heteroaryl or optionally substituted heterocyclyl;

R ^3B can be selected from O ^- OH, an optionally substituted N-linked amino acid or an optionally substituted N-linked amino acid ester;

R ^3C is an alkaneRadicals, alkenyl radicals, alkynyl radicals, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) -O- (C) ₀ -C ₂ ) (heteroaryl), -S-alkyl, -S-alkenyl, -S-alkynyl, -S- (C) ₀ -C ₂ ) (cycloalkyl), -S- (C) ₀ -C ₂ ) (heterocyclyl), -S- (C) ₀ -C ₂ ) (aryl), or-S- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

R ^3D is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) or-O- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

R ⁵ is C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ )、C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl), -OR ²⁵ 、-C(O)R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ Each of which may be optionally substituted;

R ⁶ is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the Or (b)

R ⁵ And R is ⁶ Together with the nitrogen to which they are bound may form a heterocyclic ring;

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl;

R ²² cl, br, F, CN, N of a shape of Cl, br, F, CN, N ₃ 、C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl); ONHC (=o) OR ²³ 、-NHOR ²⁴ 、-OR ²⁵ 、-SR ²⁵ 、-NH(CH ₂ ) _1-4 N(R ²⁶ ) ₂ 、-NHNHR ²⁶ 、-N＝NR ²⁷ 、-NHC(O)NHNHR ²⁷ 、-NHC(S)NHNHR ²⁷ 、-C(O)NHNHR ²⁷ 、-NR ²⁷ SO ₂ R ²⁸ 、-SO ₂ NR ²⁷ R ²⁹ 、-C(O)NR ²⁷ R ²⁹ 、-CO ₂ R ²⁹ 、-SO ₂ R ²⁹ 、-P(O)H(OR ²⁹ )、-P(O)(OR ²⁹ )(OR ³⁰ )、-P(O)(OR ²⁹ )(NR ²⁹ R ³⁰ ) or-NR ⁵ R ⁶ ；

For example, including but not limited to the following embodiments: chloro, bromo, fluoro, cyano, azido, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl, 1-dimethylpropyl, 2-dimethylpropyl, 3-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, ethynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, - (CH) ₂ ) -cyclopropyl, - (CH) ₂ ) -cyclobutyl, - (CH) ₂ ) Cyclopentyl, - (CH) ₂ ) Cyclohexyl, aziridine, oxirane (oxalane), thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene (thioplane), pyrazolidine, piperidine, oxetane (oxalane), thiane, - (CH) ₂ ) Aziridine, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Azetidine, - (CH) ₂ ) Oxetane, - (CH) ₂ ) ThietanesAlkane, - (CH) ₂ ) Pyrrolidine, - (CH) ₂ ) Tetrahydrofuran, - (CH) ₂ ) Tetrahydrothiophene, - (CH) ₂ ) Pyrazolidines, - (CH) ₂ ) Piperidine, - (CH) ₂ ) -oxacyclohexane, - (CH) ₂ ) -thiacyclohexane, phenyl, pyridinyl, -ONHC (=o) OCH ₃ 、-ONHC(＝O)OCH ₂ CH ₃ 、-NHOH、NHOCH ₃ 、-OCH ₃ 、OC ₂ H ₅ 、-OPh、OCH ₂ Ph、-SCH ₃ 、-SC ₂ H ₅ 、-SPh、SCH ₂ Ph、-NH(CH ₂ ) ₂ NH ₂ 、-NH(CH ₂ ) ₂ N(CH ₃ ) ₂ 、-NHNH ₂ 、-NHNHCH ₃ 、-N＝NH、-N＝NCH ₃ 、-N＝NCH ₂ CH ₃ 、-NHC(O)NHNH ₂ 、-NHC(S)NHNH ₂ 、-C(O)NHNH ₂ 、-NHSO ₂ CH ₃ 、-NHSO ₂ CH ₂ CH ₃ 、-SO ₂ NHCH ₃ 、-SO ₂ N(CH ₃ ) ₂ 、-C(O)NH ₂ 、-C(O)NHCH ₃ 、-C(O)N(CH ₃ ) ₂ 、-CO ₂ CH ₃ 、-CO ₂ CH ₂ CH ₃ 、-CO ₂ Ph、-CO ₂ CH ₂ Ph、-SO ₂ CH ₃ 、-SO ₂ CH ₂ CH ₃ 、-SO ₂ Ph、-SO ₂ CH ₂ Ph、-P(O)H(OH)、-P(O)H(OCH ₃ )、-P(O)(OH)(OH)、-P(O)(OH)(OCH ₃ )、-P(O)(OCH ₃ )(OCH ₃ )、-P(O)(OH)(NH ₂ )、-P(O)(OH)(NHCH ₃ )、-P(O)(OH)N(CH ₃ ) ₂ 、-NHC(O)CH ₃ 、-NHC(O)CH ₂ CH ₃ 、-NHC(O)CH(CH ₃ ) ₂ 、-NHC(O)OCH ₃ 、-NHC(O)OCH ₂ CH ₃ 、-NHC(O)OCH(CH ₃ ) ₂ 、-NHC(O)OCH ₂ CH ₂ CH ₃ 、-NHC(O)OCH ₂ CH ₂ CH ₂ CH ₃ and-NHC (O) OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ；

R ²³ Is C ₁ -C ₅ Alkyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle) - (C _0-2 Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted;

R ²⁴ is hydrogen, C ₁ -C ₆ Alkyl, - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen;

R ²⁵ is hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen;

R ²⁶ independently selected from hydrogen, C ₁ -C ₆ Alkyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen;

R ²⁷ Is hydrogen or optionally substituted C ₁ -C ₆ An alkyl group;

R ²⁸ is C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) _0- C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted;

R ²⁹ is hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) _0- C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen; or (b)

R ²⁷ And R is ²⁹ Together with the nitrogen to which they are bound may form a heterocyclic ring;

R ³⁰ is hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) _0- C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen; or (b)

R ²⁹ And R is ³⁰ May be taken together to form a heterocycle;

x is 1, 2 or 3.

beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ Metabolism of-dimethyl-2, 6-diaminopurine nucleotide includes β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ -the formation of dimethyl-2, 6-diaminopurine nucleoside triphosphates and the production of the corresponding guanine nucleoside triphosphates. See schemes 2 and 3..

2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-N ⁶ The substituted-2, 6-diaminopurine nucleotide may be in the N ² Further substitution at the position by alkylation or acylation, which may improve lipophilicity, pharmacokinetics and/or target the nucleotide to the liver. It has been found that in vitro and in vivo, in the 2-position of diaminopurinesModified 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-N ⁶ The substituted-2, 6-diaminopurine nucleotides may be dealkylated or deacylated by liver enzymes to further increase the in vivo and in vitro specificity of the nucleotide derivative, unless N ² Amino groups are completely substituted by different moieties as described herein, for example fluorine. For example, when cultured in vitro with human liver S9 fraction for up to 60 minutes (these conditions mimic in vivo conditions), nucleoside phosphoramidate 2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ² -methyl-N ⁶ Dealkylation of methyl-2, 6-diaminopurine nucleoside phosphoramidates to 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -methyl-2, 6-diaminopurine nucleoside phosphoramidate. In one embodiment, N ² Modification will increase cell permeability and liver targeting.

Despite the vast number of antiviral nucleoside literature and patent applications, 2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N as described herein has not been disclosed ⁶ -methyl-2, 6-diaminopurine nucleosides, 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -dimethyl-2, 6-diaminopurine nucleosides and other beta-D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N ⁶ 5' -stable phosphate derivatives of substituted purine nucleoside derivatives, nor are their advantageous activities described.

Unless otherwise indicated, the compounds described herein are provided in the β -D-configuration. Likewise, when in the form of a phosphoramide or phosphorothioate, the amino acid moiety may be in the L-or D-configuration. In another embodiment, the compounds may be provided in the β -L configuration. Likewise, any substituent exhibiting chirality may be provided in racemic, enantiomeric, diastereomeric forms, or any mixture thereof. When phosphoramidates, phosphorothioates, or other stable phosphorus prodrugs wherein phosphorus exhibits chirality are used as R ⁴ When a phosphate prodrug is stabilized, it may be provided as an R or S chiral phosphorus derivative or a mixture thereof (including a racemic mixture). All such combinations of spatial configurations are included in the invention described herein.

Accordingly, the present invention includes compounds of formulas I-VII, or pharmaceutically acceptable compositions, salts or prodrugs thereof, as described herein:

In a particular embodiment, the parent nucleoside, i.e., wherein R ⁴ Nucleosides that are hydrogen and thus have a hydroxyl group at the 5' -position are substantially not deaminated by adenosine deaminase in a period of 7, 10, 30, 60 or 120 minutes in a simulated in vivo environment (e.g., ambient temperature and aqueous physiological pH). Unless otherwise stated, the period of time is 30 minutes. In this embodiment, the term "substantially free of deamination" means that the parent compound is not converted to the corresponding guanine derivative or 6-oxo derivative in an amount sufficient to provide an in vivo therapeutic effect.

The compounds, methods, and compositions are provided for treating a host infected with HCV virus by administering an effective amount of the compound, or a pharmaceutically acceptable salt thereof.

The compounds and compositions may also be used to treat related disorders, such as anti-HCV antibody positive and antigen positive disorders, viral-based chronic hepatitis, liver cancer caused by advanced hepatitis c, cirrhosis, chronic or acute hepatitis c, fulminant hepatitis c, chronic persistent hepatitis c, and anti-HCV-based fatigue. The compounds or formulations comprising the compounds may also be used prophylactically to prevent or limit the progression of an anti-HCV antibody or antigen positive or clinical condition in an individual who has been exposed to hepatitis c.

In another embodiment, compounds of formula Ia are disclosed:

wherein:

Y、R ³ and R is ⁴ As defined above.

In one embodiment of formula Ia, R ³ Is hydrogen.

In one embodiment of formula Ia, when Y is NR ¹ R ² When R is ¹ Is methyl and R ² Is hydrogen.

In one embodiment of formula Ia, when Y is NR ¹ R ² When R is ¹ And R is ² Are all methyl groups.

In one embodiment of formula Ia, when Y is NR ¹ R ² When R is ¹ Is methyl and R ² Is cyclopropyl.

In another embodiment, compounds of formula Ib are disclosed

Wherein:

Y、R ³ and R is ⁴ As defined above.

In one embodiment of formula Ib, R ³ Is hydrogen.

In one embodiment of formula Ib, when Y is NR ¹ R ² When R is ¹ Methyl and R2 is hydrogen.

In one embodiment of formula Ib, when Y is NR ¹ R ² When R is ¹ And R is ² Are all methyl groups.

In one embodiment, compounds of formula II are disclosed:

wherein:

Y、R ³ 、R ⁴ 、R ¹² and R is ²² As defined above.

In another embodiment, compounds of formula IIa are disclosed:

wherein the ratio is

Y、R ³ 、R ⁴ And R is ²² As defined above.

In another embodiment, compounds of formula IIb are disclosed

Wherein:

Y、R ³ 、R ⁴ and R is ²² As defined above.

In one embodiment, compounds of formula III are disclosed

Wherein the variables Y, R ³ 、R ⁷ 、R ⁸ 、R ^9a 、R ^9b 、R ¹⁰ 、R ¹² And R is ²² As described herein.

In one embodiment, compounds of formula IV are disclosed

Wherein the variables Y, R ³ 、R ⁷ 、R ⁸ 、R ^9a 、R ^9b 、R ¹⁰ And R is ²² As described herein.

In one embodiment, compounds of formula V are disclosed

In one embodiment, compounds of formula VI are disclosed: :

wherein:

R ⁴¹ halogen (especially F OR Cl), OR ³ 、N ₃ 、NH ₂ Or CN; and

variable Y, R ³ 、R ⁴ And R is ¹² As described herein.

In one embodiment, compounds of formula VII are disclosed: :

wherein the variables Y, R ³ 、R ⁴ 、R ¹² And R is ⁴¹ As described herein.

The phosphorus in any of the above formulas may be chiral and thus may be provided as the R or S enantiomer or mixtures thereof (including racemic mixtures).

Compound 5 was separated into enantiomer compounds 5-1 and 5-2. Compound 5-2 was also prepared by chiral synthesis designated compound 24.

In one embodiment, the compounds, methods, and compositions provide a host for treating infection or exposure to hepatitis c described herein. The compounds of the present invention may be administered alone or in combination with additional anti-HCV agents in an amount effective to treat an infected host. In some embodiments, it is useful to administer a combination of drugs that modulate the same or different pathways or inhibit different targets in the virus. Due to the disclosed beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N ⁶ Substituted purine nucleotides are NS5B polymerase inhibitors, thus contacting the compound with a protease inhibitor, such as an NS3/4A protease inhibitor (e.g., telaprevirBessepir (Victrelis) ^TM ) Cimicavir(Olysio ^TM ) Or paritaprevir or NS5A inhibitors (e.g. Ombitasvir) may be useful for administration in combination to a host. The compounds of the invention may also be structurally different from NS5B polymerase inhibitors such as another compound herein or as described below (including Gilead's +.>) And (3) combined administration. The compounds of the invention may also be administered in combination with interferon alpha-2 a (which may be pegylated or otherwise modified) and/or ribavirin.

The beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N of the invention ⁶ The substituted purine nucleotides are typically administered orally, e.g. in the form of pills or tablets, but may be administered via other routes deemed appropriate by the attending physician, including via intravenous, transdermal, subcutaneous, topical, parenteral or other suitable routes.

Brief Description of Drawings

Fig. 1 is a sample chromatogram showing a semi-preparative run for separating stereoisomers of compound 5 using a Phenominex Luna column as disclosed in example 9. The y-axis is shown in mAU and the x-axis is measured in minutes. FIG. 2 is a graph of HCV replication inhibition with Compound 5-2 (Table 7) and Sofosbuvir. Compound 5-2 has EC ₅₀ ＝4nM，TC ₅₀ Greater than one hundred micromolar and a therapeutic index greater than 25,000. Sofosbuvir with EC ₅₀ ＝53nM，TC ₅₀ Greater than one hundred micromoles and a therapeutic index greater than 1,920. The y-axis is the percent of viral control and the x-axis is drug concentration (μm).

Fig. 3 is a graph of HCV replication inhibition by compound 25 (table 7) and sofosbuvir. Compound 25 has EC as described in example 27 ₅₀ ＝4nM，TC ₅₀ Greater than 100 μm and a therapeutic index greater than 25,000. Sofosbuvir with EC ₅₀ ＝53nM，TC ₅₀ Greater than one hundred micromoles and a therapeutic index greater than 1,920. The y-axis is the percent of viral control and the x-axis is drug concentration (μm).

FIG. 4 is a comparison of the anti-HCV activity of compounds 5-2, 25, 27 (Table 7) and Sofosbuvir in an intra-batch (intra-assay) assay. The y-axis is the percent of viral control and the x-axis is drug concentration (μm). See, example 27.

FIG. 5 is a diagram showing Compound 5-2; n of Compound 5-2 ² Acetic acid ester, N of Compound 5-2 ² -butyrate; n of Compound 5-2 ² -a methyl derivative; and N of Compound 5-2 ² -graph of stability of n-amyl carbamate in human blood. The x-axis is the measured incubation time (minutes) and the y-axis is a measure of the percentage of the remaining parent compound.

FIG. 6 is a graph showing 2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N in the presence of a human liver S9 fraction ² -methyl-N ⁶ -methyl-2, 6-diaminopurine nucleoside phosphoramidate to 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -in vitro time course dealkylation profile of methyl-2, 6-diaminopurine nucleoside phosphoramidates. The x-axis is the measured incubation time (minutes) and the y-axis is the concentration (nM) measurement of the remaining compounds.

FIG. 7 is a graph showing compound 5-2 in the presence of a human liver S9 fraction; n of Compound 5-2 ² Acetic acid ester, N of Compound 5-2 ² -butyrate; n of Compound 5-2 ² -a methyl derivative; and N of Compound 5-2 ² -graph of stability of n-amyl carbamate. The x-axis is the time (minutes) measured and the y-axis is a percentage measure of the remaining compounds.

Figure 8 shows the metabolites of primary compound 25 produced in human hepatocytes. The x-axis is incubation time (hours). The y-axis is the intracellular concentration (pmol/10) ⁶ Cells). See example 33..

Fig. 9 shows the metabolites of the primary compound 27 produced in human hepatocytes. The x-axis is incubation time (hours). The y-axis is the intracellular concentration (pmol/106 cells). See example 33.

FIG. 10 shows the metabolites of the primary compound 5-2 produced in human hepatocytes. The x-axis is incubation time (hours). The y-axis is the intracellular concentration (pmol/10) ⁶ Cells). See example 33.

FIG. 11 is a graph showing the activation pathways of compounds 25, 27 and 5-2. As can be seen, compounds 25, 27 and 5-2 are converted to their corresponding monophosphate analogs, which are then metabolized to common MP analogs; beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-guanine monophosphate. The monophosphate is then stepwise phosphorylated to the active triphosphate β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-guanine triphosphate. See example 33.

Detailed Description

The invention disclosed herein is a compound, method and composition for treating humans and other host animals infected with or exposed to HCV virus comprising administering an effective amount of a compound of formula I-VII as described herein, or a pharmaceutically acceptable salt or prodrug thereof, optionally in a pharmaceutically acceptable carrier. The compounds of the invention have antiviral activity or are metabolized to compounds that exhibit such activity.

The compounds and compositions may also be used to treat conditions involving or present as a consequence of HCV viral exposure. For example, the active compounds are useful in the treatment of HCV antibody positive and HCV antigen positive conditions, viral-based chronic hepatitis, liver cancer caused by advanced hepatitis c, cirrhosis, acute hepatitis c, fulminant hepatitis c, chronic persistent hepatitis c, and anti-HCV-based fatigue. In one embodiment, the compound or formulation comprising the compound may also be used prophylactically to prevent or arrest the progression of a clinical condition in an individual positive for HCV antibodies or HCV antigens or who has been exposed to hepatitis c.

In particular, it has been found that β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ -methyl-2, 6-diaminopurine nucleotides and beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -dimethyl-2, 6-diaminopurine nucleotides and other beta-D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N as described below ⁶ 5' -stable phosphate prodrugs or derivatives of substituted purine nucleotides are highly potent against HCV. This is particularly surprising because the parent nucleoside β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ Methyl-2, 6-diaminopurine in replicon assays (EC ₅₀ =15.7 micromolar) shows that it is also unsuitable as a human drug due to insufficient activity, however, the stable phosphate prodrug (phosphoramidate) is inThe replicon test showed EC ₅₀ =26 nanomoles, with an increase in activity of more than 870 fold. Likewise, the parent nucleoside β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N ⁶ -dimethyl-2, 6-diaminopurine in replicon assays (EC ₅₀ Activity in =10.7 micromolar, "μΜ") suggests that it is also unsuitable as a human drug due to insufficient activity, however, stable phosphate prodrugs (phosphoramidates) show EC in replicon assays ₅₀ =12 nanomole ("nM"), its activity increased by more than 1,300 fold.

Despite the extensive antiviral nucleoside literature and patent applications, 2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-N is not specifically disclosed ⁶ -methyl-2, 6-diaminopurine nucleotide, 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -dimethyl-2, 6-diaminopurine nucleotides and other beta-D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N ⁶ -5' -stable phosphate derivatives of substituted purine nucleotides.

Unless otherwise indicated, the compounds described herein are provided in the β -D-configuration. In another embodiment, the compounds may be provided in the β -L configuration. Likewise, any substituent exhibiting chirality may be provided in racemic, enantiomeric, diastereomeric forms, or any mixture thereof. When phosphoramidates, phosphorothioates, or other stable phosphorus prodrugs wherein phosphorus exhibits chirality are used as R ⁴ When a phosphate prodrug is stabilized, it may be provided as an R or S chiral phosphorus derivative or a mixture thereof (including a racemic mixture). The amino acid of the phosphoramidate or phosphorothioate may be in the D-or L-configuration or mixtures thereof, including racemic mixtures. All such combinations of spatial configurations are included in the invention described herein.

The invention comprises the following characteristics of

(a) Compounds of formulae I-VII as described herein, and pharmaceutically acceptable salts and prodrugs thereof;

(b) Formulas I-VII as described herein, and pharmaceutically acceptable salts and prodrugs thereof, for use in the treatment or prevention of hepatitis c virus infection;

(c) The use of pharmaceutically acceptable salts and prodrugs of formula I-VII in the manufacture of a medicament for the treatment of hepatitis c virus infection;

(d) A method of preparing a medicament intended for therapeutic use in the treatment of a hepatitis c virus infection, characterized in that formulae I-VII as described herein are used in the preparation method;

(e) A pharmaceutical formulation comprising an effective host therapeutic amount of formulae I-VII or a pharmaceutically acceptable salt or prodrug thereof, and a pharmaceutically acceptable carrier or diluent;

(f) The substantial absence of stereoisomers of the compounds or substantial separation of other chemical entities, as described herein for formulas I-VII; and

(g) A process for preparing a therapeutic product comprising an effective amount of formulas I-VII as described herein.

I. The 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N of the invention ⁶ -substituted purine nucleotides

The active compounds of the present invention are those described in formula I, which may be provided in a pharmaceutically acceptable composition, salts or prodrugs thereof:

Wherein the ratio is

Y is NR ¹ R ² ；

R ¹ Is C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CH ₂ F、CH ₂ F、CF ₃ 、CH ₂ CF ₃ 、CF ₂ CH ₃ And CF (compact F) ₂ CF ₃ )、C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl), -OR ²⁵ 、-C(O)R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ Each of which may be optionally substituted;

R ² is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CH ₂ F、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the And

R ^3B can be selected from O ^- OH, optionallyA substituted N-linked amino acid or an optionally substituted N-linked amino acid ester;

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl.

A stable phosphate prodrug is any moiety that can deliver mono-, di-, or triphosphates.

In another embodiment, compounds of formula Ia are disclosed

Wherein:

Y、R ³ and R is ⁴ As defined above.

In another embodiment, compounds of formula Ib are disclosed

Wherein:

Y、R ³ and R is ⁴ As defined above.

In another embodiment, the compound is a compound according to formula Ic

Wherein:

R ⁷ is hydrogen, C _1-6 An alkyl group; c (C) _3-7 Cycloalkyl; heteroaryl, heterocycle, or aryl, including but not limited to phenyl or naphthyl, wherein the phenyl or naphthyl is optionally substituted with: c (C) _1-6 Alkyl, C _2-6 Alkenyl, C _2-6 Alkynyl, C _1-6 Alkoxy, F, cl, br, I, nitro, cyano, C _1-6 Haloalkyl, -N (R) ⁷ ') ₂ 、C _1-6 Amido, NHSO ₂ C _1-6 Alkyl, -SO ₂ N(R ⁷ ') ₂ 、COR ^7" and-SO ₂ C _1-6 An alkyl group; (R) ⁷ ' independently hydrogen or C _1-6 An alkyl group; r is R ^7" is-OR ¹¹ or-N (R) ⁷ ) ₂ )；

R ⁸ Is hydrogen, C _1-6 Alkyl, or R ^9a Or R is ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms; wherein n is 2 to 4;

R ^9a and R is ^9b Is (i) independently selected from hydrogen, C _1-6 Alkyl, cycloalkyl, - (CH) ₂ ) _c (NR ⁹ ') ₂ 、C _1-6 Hydroxyalkyl, -CH ₂ SH、-(CH ₂ ) ₂ S(O)(Me、-(CH ₂ ) ₃ NHC(＝NH)NH ₂ (lH-indol-3-yl) methyl, (1H-imidazol-4-yl) methyl, - (CH) ₂ ) _c COR ^9" Aryl and aryl (C) _1-3 Alkyl) -, which may optionally be selected from hydroxy, C _1-6 Alkyl, C _1-6 Groups substituted with alkoxy, halogen, nitro and cyano; (ii) R is R ^9a And R is ^9b Are all C _1-6 An alkyl group; (iii) R is R ^9a And R is ^9b Together are (CH) ₂ ) _r Thereby forming a spiro ring; (iv) R is R ^9a Is hydrogen, and R ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms, (v) R ^9b Is hydrogen and R ^9a And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising contiguous N and C atoms, wherein C is 1 to 6, N is 2 to 4, R is 2 to 5, and wherein R ⁹ ' independently hydrogen or C _1-6 Alkyl, and R ^9" is-OR ¹¹ or-N (R) ¹¹ ') ₂ )；(vi)R ^9a Is hydrogen and R ^9b Is hydrogen, CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH or lower cycloalkyl; or (vii) R ^9a Is CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower cycloalkyl, and R ^9b Is hydrogen;

R ¹⁰ is hydrogen, C optionally substituted by alkoxy, di (lower alkyl) -amino or halogen _1-6 Alkyl, C _1-6 Haloalkyl, C _3-7 Cycloalkyl, heterocycloalkyl, aminoacyl, aryl, for example phenyl, heteroaryl, for example pyridinyl, substituted aryl or substituted heteroaryl;

R ¹¹ is optionally substituted C _1-6 Alkyl, optionally substituted cycloalkyl; optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl or optionally substituted acyl groups including, but not limited to, C (O) (C _1-6 An alkyl group); and

Y、R ³ and R is ¹² As defined herein.

In one embodiment, compounds of formula II are disclosed

Wherein:

y is NR ¹ R ² ；

R ² is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ CF ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the And

R ^3C is alkyl groupAlkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) -O- (C) ₀ -C ₂ ) (heteroaryl), -S-alkyl, -S-alkenyl, -S-alkynyl, -S- (C) ₀ -C ₂ ) (cycloalkyl), -S- (C) ₀ -C ₂ ) (heterocyclyl), -S- (C) ₀ -C ₂ ) (aryl) or-S- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

R ⁶ is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CH ₂ F、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ And C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the Or (b)

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl;

R ²² cl, br, F, CN, N of a shape of Cl, br, F, CN, N ₃ 、C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) _0- C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl); ONHC (=o) OR ²³ 、-NHOR ²⁴ 、-OR ²⁵ 、-SR ²⁵ 、-NH(CH ₂ ) _1-4 N(R ²⁶ ) ₂ 、-NHNHR ²⁶ 、-N＝NR ²⁷ 、-NHC(O)NHNHR ²⁷ 、-NHC(S)NHNHR ²⁷ 、-C(O)NHNHR ²⁷ 、-NR ²⁷ SO ₂ R ²⁸ 、-SO ₂ NR ²⁷ R ²⁹ 、-C(O)NR ²⁷ R ²⁹ 、-CO ₂ R ²⁹ 、-SO ₂ R ²⁹ 、-P(O)H(OR ²⁹ )、-P(O)(OR ²⁹ )(OR ³⁰ )、-P(O)(OR ²⁹ )(NR ²⁹ R ³⁰ ) or-NR ⁵ R ⁶ ；

For example, including but not limited to the following embodiments: chloro, bromo, fluoro, cyano, azido, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl, 1-dimethylpropyl, 2-dimethylpropyl, 3-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, ethynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, - (CH) ₂ ) -cyclopropyl, - (CH) ₂ ) -cyclobutyl, - (CH) ₂ ) Cyclopentyl, - (CH) ₂ ) -cyclohexyl, aziridine, oxirane, ethylthiirane (thiairane), azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene (thialane), pyrazolidine, piperidine, oxetane, thiane (thiane), - (CH) ₂ ) Aziridine, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Azetidine, - (CH) ₂ ) Oxetane, - (CH) ₂ ) Thietane, - (CH) ₂ ) Pyrrolidine, - (CH) ₂ ) Tetrahydrofuran, - (CH) ₂ ) Tetrahydrothiophene, - (CH) ₂ ) Pyrazolidines, - (CH) ₂ ) Piperidine, - (CH) ₂ ) -oxacyclohexane, - (CH) ₂ ) -thiacyclohexane, phenyl, pyridinyl, -ONHC (=o) OCH ₃ 、-ONHC(＝O)OCH ₂ CH ₃ 、-NHOH、NHOCH ₃ 、-OCH ₃ 、OC ₂ H ₅ 、-OPh、OCH ₂ Ph、-SCH ₃ 、-SC ₂ H ₅ 、-SPh、SCH ₂ Ph、-NH(CH ₂ ) ₂ NH ₂ 、-NH(CH ₂ ) ₂ N(CH ₃ ) ₂ 、-NHNH ₂ 、-NHNHCH ₃ 、-N＝NH、-N＝NCH ₃ 、-N＝NCH ₂ CH ₃ 、-NHC(O)NHNH ₂ 、-NHC(S)NHNH ₂ 、-C(O)NHNH ₂ 、-NHSO ₂ CH ₃ 、-NHSO ₂ CH ₂ CH ₃ 、-SO ₂ NHCH ₃ 、-SO ₂ N(CH ₃ ) ₂ 、-C(O)NH ₂ 、-C(O)NHCH ₃ 、-C(O)N(CH ₃ ) ₂ 、-CO ₂ CH ₃ 、-CO ₂ CH ₂ CH ₃ 、-CO ₂ Ph、-CO ₂ CH ₂ Ph、-SO ₂ CH ₃ 、-SO ₂ CH ₂ CH ₃ 、-SO ₂ Ph、-SO ₂ CH ₂ Ph、-P(O)H(OH)、-P(O)H(OCH ₃ )、-P(O)(OH)(OH)、-P(O)(OH)(OCH ₃ )、-P(O)(OCH ₃ )(OCH ₃ )、-P(O)(OH)(NH ₂ )、-P(O)(OH)(NHCH ₃ )、-P(O)(OH)N(CH ₃ ) ₂ 、-NHC(O)CH ₃ 、-NHC(O)CH ₂ CH ₃ 、-NHC(O)CH(CH ₃ ) ₂ 、-NHC(O)OCH ₃ 、-NHC(O)OCH ₂ CH ₃ 、-NHC(O)OCH(CH ₃ ) ₂ 、-NHC(O)OCH ₂ CH ₂ CH ₃ 、-NHC(O)OCH ₂ CH ₂ CH ₂ CH ₃ and-NHC (O) OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ；

R ²⁷ hydrogen or optionally substituted C ₁ -C ₆ An alkyl group;

R ²⁹ And R is ³⁰ May be taken together to form a heterocycle;

x is 1, 2 or 3.

In another embodiment, compounds of formula IIa are disclosed:

wherein:

Y、R ³ 、R ⁴ and R is ²² As defined above.

In another embodiment, compounds of formula IIb are disclosed

Wherein:

Y、R ³ 、R ⁴ and R is ²² As defined above.

In a typical embodiment, the compound is the β -D isomer relative to the corresponding nucleoside (i.e., naturally occurring configuration). In an alternative configuration, the compound is provided as the β -L isomer. The compounds are generally at least 90% free of the opposite enantiomer, and may be at least 98%, 99% or even 100% free of the opposite enantiomer. Unless otherwise stated, the compounds are at least 90% free of the opposite enantiomer.

In another embodiment, the compound is a compound according to formula III

Wherein:

R ^9a and R is ^9b Is (i) independently selected from hydrogen, C _1-6 Alkyl, cycloalkyl, - (CH) ₂ ) _c (NR ⁹ ') ₂ 、C _1-6 Hydroxyalkyl, -CH ₂ SH、-(CH ₂ ) ₂ S(O)Me、-(CH ₂ ) ₃ NHC(＝NH)NH ₂ (lH-indol-3-yl) methyl, (1H-imidazol-4-yl) methyl, - (CH) ₂ ) _c COR ^9" Aryl and aryl (C) _1-3 Alkyl) -, which aryl group may be optionally substituted with a group selected from: hydroxy, C _1-6 Alkyl, C _1-6 Alkoxy, halogen, nitro and cyano; (ii) R is R ^9a And R is ^9b Taken together as C _1-6 An alkyl group; (iii) R is R ^9a And R is ^9b Together are (CH) ₂ ) _r Thereby forming a spiro ring; (iv) R is R ^9a Is hydrogen and R ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms, (v) R ^9b Is hydrogen and R ^9a And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising contiguous N and C atoms, wherein C is 1 to 6, N is 2 to 4, R is 2 to 5, and wherein R ⁹ ' independently hydrogen or C _1-6 Alkyl, and R ⁹ "is-OR ¹¹ or-N (R) ¹¹ ') ₂ )；(vi)R ^9a Is hydrogen and R ^9b Is hydrogen, CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower cycloalkyl; or (vii) R ^9a Is CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower alkyl and R ^9b Is hydrogen;

R ¹⁰ is hydrogen, C _1-6 Is hydrogen, C optionally substituted by alkoxy, di (lower alkyl) -amino or halogen _1-6 Alkyl, C _1-6 Haloalkyl, C _3-7 Cycloalkyl, heterocycloalkyl, aminoacyl, aryl, for example phenyl, heteroaryl, for example pyridinyl, substituted aryl or substituted heteroaryl;

Y、R ³ 、R ¹² and R is ²² As defined above.

In one embodiment, compounds of formula IV are disclosed:

In one embodiment, compounds of formula V are disclosed

In another embodiment, the compounds, methods, and compositions provide a host for treating infection or exposure to hepatitis c.

In one embodiment, compounds of formula VI are disclosed:

wherein:

R ⁴¹ halogen (especially F OR Cl), OR ³ (including OH, N) ₃ 、NH ₂ Or CN; and

variable Y, R ³ 、R ⁴ And R is ¹² As described herein.

In one embodiment, compounds of formula VII are disclosed:

beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-C-substituted-N ⁶ -metabolism of substituted-2, 6-diaminopurine nucleotides

beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ Metabolism of methyl-2, 6-diaminopurine nucleoside phosphoramidates involves the production of 5' -monophosphate followed by N ⁶ -methyl-2, 6-diaminopurine base anabolism produces β -D-2 '-deoxy-2' - α -fluoro-2 '- β -methyl-guanosine, which is a 5' -monophosphate. The monophosphate is then further anabolized to an active species; 5' -triphosphates. beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methylIC with anti-HCV genotype 1b NS5B polymerase of base-guanine triphosphate ₅₀ =0.15 μΜ. beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ The metabolic pathway of the methyl-2, 6-diaminopurine nucleoside phosphoramidate is illustrated in scheme 1 below.

Scheme 1

beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ Metabolism of-dimethyl-2, 6-diaminopurine nucleotide includes formation of beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ -dimethyl-2, 6-diaminopurine nucleoside triphosphates and the corresponding guanosine triphosphates are produced. These metabolic pathways are illustrated in schemes 2 and 3 below. .

Scheme 2

Scheme 3

Stable phosphate prodrugs

Stable phosphate prodrugs are moieties that can deliver mono-, di-or tri-phosphates in vivo. For example, mcGuigan is described in U.S. Pat. Nos. 8,933,053;8,759,318;8,658,616;8,263,575;8,119,779; phosphoramidates are disclosed in 7,951,787 and 7,115,590. Alios discloses phosphorothioates in US 8,895,723 and 8,871,737 (incorporated herein by reference). The cyclic nucleotides are also disclosed in U.S. Pat. No.8,772,474 (incorporated herein by reference). Idenix discloses cyclic phosphoramidates and phosphoramidate/SATE derivatives in WO 2013/177219 (incorporated herein by reference). Idenix also discloses substituted carbonyloxymethyl phosphoramidate compounds in WO 2013/039920 (incorporated herein by reference) And (3) an object. Hostetler discloses lipid phosphate prodrugs, see for example US 7,517,858.Hostetler also discloses conjugates of phosphonate prodrugs, see for example US 8,889,658;8,846,643;8,710,030;8,309,565;8,008,308; and 7,790,703. The nucleotide sphingosine and lipid derivatives are disclosed in WO 2014/124430 by Ememory University. RFS Pharma discloses purine nucleoside monophosphate prodrugs in WO 2010/091386. Purine nucleoside monophosphate prodrugs are also disclosed in U.S. Pat. No. 5, 9,173,893, incorporated herein by reference. HepDIRECT ^TM Additional phosphate prodrugs include, but are not limited to, phosphate esters, 3',5' -cyclic phosphate esters (including cyclosaL), SATE derivatives (S-acyl-2-thio esters) and DTE (dithiodiethyl) prodrugs in the articles "Design, synthesis, and Characterization of a Series of Cytochrome P (450) 3A-Activated Prodrugs (HepDirect Prodrugs) Useful for Targeting Phosph (on) ate-Based Drugs to the Liver," (J.am. Chem. Soc.126,5154-5163 (2004), ", for reviews of references disclosing non-limiting examples, see: A.ray and K.Hostetler," Application of kinase bypass strategies to nucleoside antivirals, "Antiviral Research (2011) 277-291; M.Sofia," Nucleotide prodrugs for HCV therapy, "Antiviral Chemistry and Chemotherapy 2011;22-23-49; and S.Peyrottes et al," SATE Pronucleotide Approaches: an oview, "Mini Reviews in Medicinal Chemistry 2004,4,395. In one embodiment, the 5' -prodrugs described in any of these patent applications or documents may be used in the present invention R compounds ⁴ Bits. .

In an alternative embodiment, stable phosphate prodrugs include, but are not limited to, those described in U.S. Pat. No.9,173,893 and U.S. Pat. No.8,609,627 (incorporated herein by reference), including methods of preparation thereof. For example, the 5' -prodrugs of formulas I-V may be represented by the following groups:

in an alternative embodiment, the 3',5' -prodrug of formula I-V may be represented by the following groups:

wherein the ratio is

When chiral is present in the phosphorous (phosphous) center, it may be all or part of R _p Or S _p Or any mixture thereof.

Z is O or S;

R ³³ selected from OR ³⁴ 、And derived fatty alcohols (e.g., without limitation: linoleyl +)>Oil base->)

Wherein R is ³⁴ 、R ³⁵ And R is ³⁶ Is defined as follows;

R ³¹ and R is ³² When administered in vivo, nucleoside or thiomonophosphates can be provided which may or may not be partially or fully resistant to 6-NH in biological systems ₂ Deamination. Representative R ³¹ And R is ³² Independently selected from

(a)OR ³⁴ Wherein R is ³⁴ Selected from H, li, na, K, phenyl and pyridinyl; phenyl and pyridyl are independently selected from one to three (CH) ₂ ) _0-6 CO ₂ R ³⁷ And (CH) ₂ ) _0-6 CON(R ³⁷ ) ₂ Is substituted by a substituent of (a);

R ³⁷ h, C independently _1-20 Alkyl, carbon chain derived from fatty alcohols (e.g., oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.), or C substituted with _1-20 Alkyl: lower alkyl, alkoxy, di (lower)Alkyl) -amino, fluoro, C _3-10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl, such as phenyl, heteroaryl, such as pyridyl, substituted aryl, or substituted heteroaryl; wherein the substituents are C _1-5 Alkyl or C substituted by _1-5 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl or cycloalkyl;

(b)

(c) Esters of D-amino acids or L-amino acids

Wherein R is ³⁶ Limited to those side chains which occur in natural L-amino acids, and

R ³⁵ h, C of a shape of H, C _1-20 Alkyl, carbon chain derived from fatty alcohols (e.g., oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.), or C substituted with _1-20 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl, such as phenyl, heteroaryl, such as pyridyl, substituted aryl, or substituted heteroaryl; wherein the substituents are C _1-5 C substituted by alkyl or by _1-5 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl or cycloalkyl;

(d)R ³¹ and R is ³² Can be combined together to form a ring

Wherein R is ³⁸ H, C of a shape of H, C _1-20 Alkyl, C _1-20 Alkenyl groups, carbon chains derived from fatty alcohols (e.g., oleyl alcohol, octacosanol, triacontanol, linoleyl alcohol, etc.), or are taken up by Substituted C _1-20 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl, such as phenyl, heteroaryl, such as pyridyl, substituted aryl, or substituted heteroaryl; wherein the substituents are C _1-5 Alkyl, or C substituted by _1-5 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl or cycloalkyl;

(e)R ³¹ and R is ³² May be joined together to form a ring selected from the group consisting of:

wherein R is ³⁹ Is O or NH, and

R ⁴⁰ selected from H, C _1-20 Alkyl, C _1-20 Alkenyl, carbon chain derived from fatty acid (e.g., oleic acid, linoleic acid, etc.), and C substituted with _1-20 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl, cycloalkylalkyl, cycloheteroalkyl, aryl, such as phenyl, heteroaryl, such as pyridyl, substituted aryl, or substituted heteroaryl; wherein the substituents are C _1-5 Alkyl or C substituted by _1-5 Alkyl: lower alkyl, alkoxy, di (lower alkyl) -amino, fluoro, C _3-10 Cycloalkyl or cycloalkyl.

The compounds may be prepared, for example, by preparing 5' -OH analogs and then converting these compounds to monophosphate analogs.

Description of the embodiments

Particular embodiments:

(i) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(ii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(iii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(iv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(v) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(vi) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(vii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(viii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(ix) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ A stable phosphorothioate prodrug;

(x) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xi) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xiii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(xiv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(xv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(xvi) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(xvii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xviii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xix) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xx) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is methyl, and R ⁴ Is a diphosphate;

(xxi) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(xxii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(xxiii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(xxiv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xxv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xxvi) In formula Ia, Y is NR ¹ R ² ,R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xxvii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(xxviii) In formula Ia, Y is NR ¹ R ² Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is propyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(xxix) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(xxx) In formula Ia, Y is NR ¹ R ² ,R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(xxxi) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xxxii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xxxiii) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xxxiv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(xxxv) In formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is ethyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(xxxvi) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(xxxvii) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(xxxviii) In formula Ib, Y is NR ¹ R ² ,R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xxxix) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xl) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xli) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(xlii) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(xliii) In formula IbY is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(xliv) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(xlv) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(xlvi) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(xlvii) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(xlviii) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a diphosphate;

(xlix) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen, R ³ Is hydrogen, and R ⁴ Is a triphosphate;

(l) In formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, R ⁴ Is a stable phosphate prodrug;

(li) in formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a stable phosphorothioate prodrug;

(lii) in formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is phosphoramidate;

(liii) in formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is phosphorothioate:

(liv) in formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a monophosphate ester;

(lv) in formula Ib, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is methyl, and R ⁴ Is a diphosphate;

(lvi) in formula Ia, Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, and R ⁴ Is a triphosphate.

In an alternative embodiment of any of the foregoing, the compound has R ²² A substituent. In certain of these particular embodiments, R ²² Is F, an amide or a carbamate. In other particular aspects of the above embodiments, R ²² Is chloro, bromo, cyano, azido, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl, 1-dimethylpropyl, 2-dimethylpropyl, 3-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, ethynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, - (CH) ₂ ) -cyclopropyl, - (CH 2) -cyclobutyl, - (CH 2) -cyclopentyl, - (CH ₂ ) -cyclohexyl, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, pyrazolidine, piperidine, oxetane, thiane, - (CH) ₂ ) Aziridine, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Azetidine, - (CH) ₂ ) Oxetane, - (CH) ₂ ) Thietane, - (CH) ₂ ) Pyrrolidine, - (CH) ₂ ) Tetrahydrofuran, - (CH) ₂ ) Tetrahydrothiophene, - (CH) ₂ ) Pyrazolidines, - (CH) ₂ ) Piperidine, - (CH) ₂ ) -oxacyclohexane, - (CH) ₂ ) ThiocyclohexanePhenyl, pyridinyl, -ONHC (=o) OCH ₃ 、-ONHC(＝O)OCH ₂ CH ₃ 、-NHOH、NHOCH ₃ 、-OCH ₃ 、OC ₂ H ₅ 、-OPh、OCH ₂ Ph、-SCH ₃ 、-SC ₂ H ₅ 、-SPh、SCH ₂ Ph、-NH(CH ₂ ) ₂ NH ₂ 、-NH(CH ₂ ) ₂ N(CH ₃ ) ₂ 、-NHNH ₂ 、-NHNHCH ₃ 、-N＝NH、-N＝NCH ₃ 、-N＝NCH ₂ CH ₃ 、-NHC(O)NHNH ₂ 、-NHC(S)NHNH ₂ 、-C(O)NHNH ₂ 、-NHSO ₂ CH ₃ 、-NHSO ₂ CH ₂ CH ₃ 、-SO ₂ NHCH ₃ 、-SO ₂ N(CH ₃ ) ₂ 、-C(O)NH ₂ 、-C(O)NHCH ₃ 、-C(O)N(CH ₃ ) ₂ 、-CO ₂ CH ₃ 、-CO ₂ CH ₂ CH ₃ 、-CO ₂ Ph、CO ₂ CH ₂ Ph、-SO ₂ CH ₃ 、-SO ₂ CH ₂ CH ₃ 、-SO ₂ Ph、-SO ₂ CH ₂ Ph、-P(O)H(OH)、-P(O)H(OCH ₃ )、-P(O)(OH)(OH)、-P(O)(OH)(OCH ₃ )、-P(O)(OCH ₃ )(OCH ₃ )、-P(O)(OH)(NH ₂ )、-P(O)(OH)(NHCH ₃ )、-P(O)(OH)N(CH ₃ ) ₂ 、-NHC(O)CH ₃ 、-NHC(O)CH ₂ CH ₃ 、-NHC(O)CH(CH ₃ ) ₂ 、-NHC(O)OCH ₃ 、-NHC(O)OCH ₂ CH ₃ 、-NHC(O)OCH(CH ₃ ) ₂ 、-NHC(O)OCH ₂ CH ₂ CH ₃ 、-NHC(O)OCH ₂ CH ₂ CH ₂ CH ₃ and-NHC (O) OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ；

In an alternative embodiment of compounds (I) to (lvi), L-nucleosides are used in formulas I-VII.

In an alternative embodiment, formula (la)R in I ¹² The variable being CH ₂ F。

In an alternative embodiment, R in formula I ¹² The variable being CHF ₂ 。

In an alternative embodiment, R in formula I ¹² The variable being CF ₃ 。

In one embodiment, compounds of formula Ia are provided. Non-limiting examples of compounds of formula Ia include:

in one embodiment, phosphorothioate of formula Ia is provided. Non-limiting examples of phosphorothioates of formula Ia include, but are not limited to

In one embodiment, stable phosphate prodrugs of formula Ia are provided. Non-limiting examples of stable phosphate prodrugs of formula Ia are illustrated below:

in another embodiment, compounds of formula Ia are provided. Non-limiting examples of compounds of formula Ia include

In one embodiment, stable phosphate prodrugs of formula Ia are provided. Non-limiting examples of stable phosphate prodrugs of formula Ia are illustrated below: :

in one embodiment, compounds of formula II are provided. Non-limiting examples of compounds of formula II include

In one embodiment, compounds of formula I are provided. Non-limiting examples of compounds of formula I include:

In one embodiment, R ⁴ Is that

In certain embodiments, R ³ Is H and R ⁴ Is that

In one embodiment, compounds of formula II are provided. Non-limiting examples of compounds of formula II include:

in certain embodiments, R ³ Is H and R ⁴ Is that

In certain embodiments, R ³ Is H and R ⁴ Is that

In certain embodiments, R ¹ Is CH ₃ ，R ² Is H, R ³ Is H and R ⁴ Is that

In certain embodiments, R ¹ Is CH ₃ ，R ² Is CH ₃ ，R ³ Is H and R ⁴ Is that

In certain embodiments, R ¹ Is cyclopropyl, R ² Is CH ₃ ，R ³ Is H and R ⁴ Is that

II. Definition of

The invention will be described using the following terminology. Where a term is not specifically defined herein, that term is given its art-recognized meaning by one of ordinary skill in the art in view of the description of the invention in this context.

The term "alkyl" shall in this context refer to a straight or branched chain fully saturated hydrocarbon or alkyl group, which may optionally be substituted (e.g. by halogen (including F)). For example, the alkyl group may have 1, 2,3, 4, 5, 6, 7, or 8 carbon atoms (i.e., C ₁ -C ₈ Alkyl), 1, 2,3, 4, 5, or 6 carbon atoms (i.e., C ₁ -C ₆ Alkyl) or 1 to 4 carbon atoms (i.e., C ₁ -C ₄ Alkyl). Examples of suitable alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, neopentyl, hexyl, 2-methylpentyl, 3-methylpentyl, 2-dimethylbutyl and 2, 3-dimethylbutyl.

The term "alkenyl" refers to non-aromatic hydrocarbon groups containing at least one double bond between adjacent carbon atoms and structures similar to alkyl groups as otherwise described herein. For example, the alkenyl group may have 2 to 8 carbon atoms (i.e., C ₂ -C ₈ Alkenyl), or 2 to 4 carbon atoms (i.e., C ₂ -C ₄ Alkenyl). Examples of suitable alkenyl groups include, but are not limited to, vinyl (ethyl) or vinyl (vinyl) (-ch=ch) ₂ ) Allyl (-CH) ₂ CH＝CH ₂ ) 1-butenyl (-c=ch-CH) ₂ CH ₃ ) And 2-butenyl (-CH) ₂ CH＝CHCH ₂ ). The alkenyl groups may be optionally substituted as described herein.

The term "alkynyl" refers to non-aromatic hydrocarbon groups containing at least one triple bond between adjacent carbon atoms and structures similar to alkyl groups as otherwise described herein. For example, an alkynyl group may have 2 to 8 carbon atoms (i.e., a C2-C8 alkyne), or 2 to 4 carbon atoms (i.e., C ₂ -C ₄ Alkynyl). Examples of alkynyl groups include, but are not limited to, acetylene (vinyl) groups or acetylene (vinyl) groups and propargyl groups. Alkynyl groups may be optionally substituted as described herein.

The term "acyl" refers to the-C (O) R moietyA moiety wherein the carbonyl moiety is bonded to R, e.g., -C (O) alkyl. R may be selected from alkoxy, alkyl, cycloalkyl, lower alkyl (i.e., C ₁ -C ₄ ) The method comprises the steps of carrying out a first treatment on the surface of the Alkoxyalkyl groups, including methoxymethyl; aralkyl groups, including benzyl; aryloxyalkyl-such as phenoxymethyl; aryl groups, including phenyl, optionally substituted with halogen, C ₁ To C ₄ Alkyl or C ₁ To C ₄ Alkoxy substitution. In one embodiment, the term "acyl" refers to mono-, di-, or triphosphate esters.

The term "lower acyl" refers to an acyl group in which the carbonyl moiety is lower alkyl (i.e., C ₁ -C ₄ )。

The term "alkoxy" refers to the group-OR ', wherein-OR' is-O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) or-O- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

the term "amino" refers to the group-NH ₂ 。

The term "amino acid" or "amino acid residue" refers to a D-or L-naturally or non-naturally occurring amino acid. Representative amino acids include, but are not limited to, alanine, beta-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan, tyrosine, or the like.

The term "azido" refers to the group-N ₃ 。

The term "aryl" or "aromatic" in this context refers to a substituted (as described elsewhere herein) or unsubstituted monovalent aromatic group having a single ring (e.g., phenyl or benzyl) or a fused ring (e.g., naphthyl, anthryl, phenanthryl, etc.), and may be bound to a compound according to the invention at any available stable site on the ring or at a site otherwise indicated in the chemical structure as presented. The aryl groups may be optionally substituted as described herein.

"cycloalkyl", "carbocycle" or "carbocyclyl" refers to a saturated (i.e., cycloalkyl) or partially unsaturated (e.g., cycloalkenyl, cyclodienyl, etc.) ring having 3 to 7 carbon atoms in a single ring. Monocyclic carbocycles have 3 to 7 ring atoms, still more typically 5 or 6 ring atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, l-cyclohex-1-enyl, l-cyclohex-2-enyl and 1-cyclohex-3-enyl.

The term "cyano" refers to the group-CN.

The term "halogen" or "halo" refers to chlorine, bromine, fluorine or iodine.

Heteroaryl ring systems are saturated or unsaturated rings having one or more nitrogen, oxygen, or sulfur atoms in the ring (monocyclic ring), including, but not limited to, imidazole, furyl (furanyl), pyrrole, furyl (furanyl), thiophene, thiazole, pyridine, pyrimidine, purine, pyrazine, triazole, oxazole, or fused ring systems such as indole, quinoline, etc., which may be optionally substituted as described above. Heteroaryl groups include nitrogen-containing heteroaryl groups such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, triazine, tetrazole, indole, isoindole, indolizine, purine, indazole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline (carbazoline), naphthyridine, phenanthroline, phenazine, oxadiazole, benzimidazole, pyrrolopyridine, pyrrolopyrimidine, and pyridopyrimidine; sulfur-containing aromatic heterocycles such as thiophene and benzothiophene; oxygen-containing aromatic heterocycles such as furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; aromatic heterocycles comprising two or more heteroatoms selected from nitrogen, sulfur and oxygen, such as thiazole, thiadiazole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, isoxazole, furazan, phenoxazine, pyrazolooxazole, imidazothiazole, thienofuran, furopyrrole, pyridooxazine, furopyridine, furopyrimidine, thienopyrimidine, oxazole and the like, all of which may be optionally substituted.

The term "heterocycle" or "heterocyclic" refers to a cyclic group containing at least one heteroatom (i.e., O, N or S) and may be aromatic (heteroaryl) or non-aromatic. Exemplary non-aromatic heterocyclic groups for use in the present invention include, for example, pyrrolidinyl, piperidinyl, piperazinyl, N-methylpiperazinyl, imidazolinyl, pyrazolidinyl, imidazolidinyl, morpholinyl, tetrahydropyranyl, azetidinyl, oxetanyl, oxathiolanyl, pyridone, 2-pyrrolidone, ethyleneurea, 1, 3-dioxolane, 1, 3-dioxane, 1, 4-dioxane, phthalimide, succinimide, and the like, all of which may be optionally substituted.

The term "hydroxy" refers to the group-OH.

The term "nitro" refers to the group-NO ₂ 。

The term "pharmaceutically acceptable salt" or "prodrug" describes throughout the specification β -D-2'- α -fluoro-2' - β -C-substituted-2-modified-N ⁶ Any pharmaceutically acceptable form of substituted purine nucleotide (e.g., ester, phosphoramidate, phosphorothioate, phosphate, salt of an ester or related group) which when administered to a patient provides the desired active compound. Examples of pharmaceutically acceptable salts are organic addition salts with acids which form physiologically acceptable anions such as tosylate, mesylate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, alpha-ketoglutarate and alpha-glycerophosphate. Suitable inorganic salts may also be formed, including sulfate, nitrate, bicarbonate, and carbonate. Pharmaceutically acceptable salts can be obtained using standard methods well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid which imparts a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts of carboxylic acids may also be prepared.

"pharmaceutically acceptable prodrugs" refers to compounds which are metabolized (e.g., hydrolyzed or oxidized) in the host to form the compounds of the invention.Typical examples of prodrugs include compounds having a biologically labile protecting group on the functional moiety of the active compound. Prodrugs include compounds that can be oxidized, reduced, aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed, dehydrated, alkylated, dealkylated, acylated, deacylated, phosphorylated, dephosphorylated, thioaminophosphorylated, desulphated aminophosphorylated, or deaminated to produce an active compound. The compounds of the present invention have antiviral activity against HCV, or are metabolized to compounds that exhibit such activity. beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N ⁶ The substituted purine nucleosides may also be administered in the form of 5' -phosphoether lipids, phosphorodiamidates, 3',5' -cyclic phosphoramidates, 3',5' -cyclic phosphorothioates, DTE conjugates, mixed phosphoramidate-SATE derivatives or "SATE" derivatives.

The term "phosphonic acid" refers to the group-P (O) (OH) ₂ 。

In one embodiment, the term purine or pyrimidine base includes, but is not limited to adenine, N ⁶ Alkylpurine, N ⁶ Acylpurines (wherein acyl is-C (O) alkyl, -C (O) (aryl) C ₀ -C ₄ Alkyl, or-C (O) (C) ₀ -C ₄ Alkyl) aryl), N ⁶ Benzyl purine, N ⁶ Halogenated purines, N ⁶ Vinyl purine, N ⁶ Alkynylpurine, N ⁶ -acylpurines, N ⁶ Hydroxy alkyl purines, N ⁶ Thioalkylpurine, N ² Alkylpurine, N ² -alkyl-6-thiopurine, thymine, cytosine, 5-fluorocytosine, 5-methylcytosine, 6-azapyrimidine (including 6-azacytosine), 2-and/or 4-mercaptopyrimidine, uracil, 5-halouracil (including 5-fluorouracil), C ⁵ -alkyl pyrimidines, C ⁵ -benzyl pyrimidine, C ⁵ -halopyrimidines, C ⁵ Vinyl pyrimidine, C ⁵ -alkynyl pyrimidines, C ⁵ -acyl pyrimidines, C ⁵ Hydroxyalkyl purines, C ⁵ -amido pyrimidines, C ⁵ -cyanopyrimidines, C ⁵ Nitropyrimidine, C ⁵ Aminopyrimidines, N ² -an alkanePurines, N ² -alkyl-6-thiopurine, 5-azacytidinyl, 5-azauridine, triazolopyridinyl, imidazopyridinyl, pyrrolopyrimidinyl and pyrazolo-pyrimidinyl. Purine bases include, but are not limited to, guanine, adenine, hypoxanthine, 2, 6-diaminopurine, and 6-chloropurine. The functional oxygen and nitrogen groups on the base may be protected as needed or desired. Suitable protecting groups are well known to those skilled in the art and include benzyl, trimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, trityl, alkyl and acyl groups such as acetyl and propionyl; methanesulfonyl and p-toluenesulfonyl. Alternatively, the purine or pyrimidine base may be optionally substituted so that it forms a useful prodrug thereof which can be cleaved in vivo. Examples of suitable substituents include acyl moieties.

The term "substituted" or "optionally substituted" refers to a moiety that may have at least one additional substituent including, but not limited to, halogen (F, cl, br, I), OH, phenyl, benzyl, N ₃ CN, acyl, alkyl including methyl; alkenyl, alkynyl, alkoxy, haloalkyl; including CHF ₂ 、CH ₂ F and CF ₃ The method comprises the steps of carrying out a first treatment on the surface of the Etc. In one embodiment, the term "substituted" or "optionally substituted" refers to a moiety that may have at least one additional substituent including, but not limited to, azido, cyano, halogen (fluoro, chloro, bromo or iodo), alkyl, alkenyl, alkynyl, cycloalkyl, heterocycle, aryl, heteroaryl, haloalkyl, hydroxy, alkoxy, amino, -NH (C) ₁ -C ₆ Unsubstituted alkyl), -NH (C) ₁ -C ₆ Substituted alkyl), -NH- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₈ Cycloalkyl), -NH- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₈ Heterocycle), -NH- (C) ₀ -C ₂ Alkyl) (aryl), -N (C) ₁ -C ₆ Unsubstituted alkyl group ₂ 、-N(C ₁ -C ₆ Unsubstituted alkyl) (C ₁ -C ₆ Substituted alkyl), -N (C) ₁ -C ₆ Substituted alkanesBase group ₂ 、-NH-(C ₀ -C ₂ Alkyl) (C) ₃ -C ₈ Cycloalkyl), -NH- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₈ Heterocycle), -NH- (C) ₀ -C ₂ Alkyl) (aryl), acyl, nitro, sulfonic acid, sulfate, phosphonic acid, phosphate, phosphonate, or thiol.

The term "sulfonate" is defined by the term "sulfonate" as including R ¹⁴ R of (2) ¹⁴ S(O) ₂ OR ¹⁵ Represented by R, wherein ¹⁴ Is alkyl, haloalkyl, aralkyl, or aryl. R is R ¹⁵ Is alkyl, aryl or aralkyl.

The term "sulfonic acid" refers to the group-SO ₂ OH。

The term "thiol" refers to the group-SH.

The term "nitrogen-protecting group" as used herein refers to a moiety covalently attached to nitrogen that can be removed and typically replaced with hydrogen when appropriate. For example, the nitrogen protecting group may be one that is removed in vivo after administration to a host, by cells in vitro, or it may be removed during the manufacturing process. Suitable nitrogen protecting groups for use in the present invention are those described by Greene and Wuts in Protective Groups in Organic Synthesis (1991) New York, john Wiley and Sons, inc.

The term "oxygen-protecting group" as used herein refers to a moiety covalently attached to oxygen that can be removed and typically replaced with hydrogen when appropriate. For example, the oxygen protecting group may be a group that is removed in vivo after administration to a host, is removed by a cell in vitro, or it may be removed during the manufacturing process. Suitable oxygen protecting groups for use in the present invention are those described by Greene and Wuts in Protective Groups in Organic Synthesis (1991) New York, john Wiley and Sons, inc.

"phosphate" refers to the group-OP (O) (OH) ₂ 。

Unless otherwise indicated, "phosphate ester" refers to mono-, di-, and tri-phosphates.

The term "phosphoramidate", "phosphoramidate" or "phosphoramidate" is a moiety having a phosphorus bound to three oxy groups and one amine (which may be optionally substituted). Suitable phosphoramidates for use in the present invention are those described by Madela, karolina and McGuigan in 2012, "Progress in the development of anti-hepatitis C virus nucleoside and nucleotide prodrugs", future Medicinal Chemistry 4 (5), pages 625-650, 10:1021/jm300074y, and Dominique, mcGuigan and Balzarini in 2004, "Aryloxy Phosphoramidate Triesters as Pro-Tides", mini Reviews in Medicinal Chemistry 4 (4), pages 371-381. Further phosphoramidates for use in the present invention are those described in U.S. Pat. No.: 5,233,031, 7,115,590, 7,547,704, 7,879,815, 7,888,330, 7,902,202, 7,951,789, 7,964,580, 8,071,568;8,148,349, 8,263,575, 8,324,179, 8,334,270, 8,552,021, 8,563,530, 8,580,765, 8,735,372, 8,759,318; EP 2120565; EP 1143995;6,455,513; and 8,334,270. Other phosphoramidates are described in the nucleoside patents described in the background of the present invention.

Phosphoramidate groups for use in the present invention include those of the following structures:

other phosphoramidate groups useful in the present invention include those of the following structures:

wherein:

R ^P1 is an optionally substituted linear, branched or cyclic alkyl group, or an optionally substituted aryl, heteroaryl or heterocyclic group or a linked combination thereof; and

R ^P2 is-NR ^N1 R ^N2 A group or B';

wherein:

R ^N1 and R is ^N2 Each independently H, C _1-8 Alkyl, (C) ₃ -C ₇ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, or (heteroaryl) C ₀ -C ₄ Alkyl-; which may be optionally substituted; or (b)

R ^N1 And R is ^N2 Taken together with the nitrogen atom to which it is attached, forms a 3 to 7 membered heterocyclic ring;

b' isA group;

wherein:

R ¹⁶ is hydrogen, (C) ₁ -C ₈ ) Alkyl, (C) ₂ -C ₈ ) Alkenyl group (C) ₂ -C ₈ ) Alkynyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, (heteroaryl) C ₀ -C ₄ Alkyl, or amino acid side chains, for example side chains typically selected from the following amino acids (as described elsewhere herein): alanine, beta-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine (typically R) ¹⁶ Hydrogen, methyl, isopropyl or isobutyl);

R ¹⁷ is hydrogen, (C) ₁ -C ₈ ) Alkyl, (C) ₂ -C ₈ ) Alkenyl group (C) ₂ -C ₈ ) Alkynyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, (heteroaryl) C ₀ -C ₄ Alkyl, or amino acid side chains, for example side chains typically selected from the following amino acids (as described elsewhere herein): alanine, beta-alanine, arginine, asparagine, aspartic acid, cysteineAmino acid, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine (typically, R ¹⁷ Hydrogen, methyl, isopropyl or isobutyl);

R ¹⁸ is hydrogen or C ₁ -C ₃ An alkyl group; or (b)

R ¹⁶ And R is ¹⁷ Can be formed into (C) ₃ -C ₇ ) Cycloalkyl or (C) ₃ -C ₇ ) A heterocyclic group; or (b)

R ¹⁸ And R is ¹⁶ Or R is ¹⁷ Can be formed into (C) ₃ -C ₆ ) A heterocyclic group; and

R ¹⁹ is hydrogen, (C) ₁ -C ₆ ) Alkyl, (C) ₃ -C ₆ ) Alkenyl group (C) ₃ -C ₆ ) Alkynyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, (heteroaryl) C ₀ -C ₄ Alkyl-; or (b)

B' isA group;

wherein:

R ²⁰ is hydrogen, (C) ₁ -C ₃ ) Alkyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, or (heteroaryl) C ₀ -C ₄ Alkyl-;

R ²¹ is hydrogen, (C) ₁ -C ₃ ) Alkyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, or (heteroaryl) C ₀ -C ₄ Alkyl-; and

R ¹⁸ and R is ¹⁹ As defined above.

Preferred R ^P1 Groups include optionally substituted phenyl, naphthyl and monocyclic heteroaryl groups, particularly those groups (particularly lipophilic groups) that increase the bioavailability of the compound in the patient's cells and exhibit reduced toxicity, increased therapeutic index and increased pharmacokinetics (slower metabolism and excretion of the compound).

The term phosphoramidate is used throughout the specification to describe groups which are present at the 5 'or 3' position of the furanose ring of a nucleoside compound and which form a prodrug form of the nucleoside compound. In one embodiment, phosphoramidates may be present at both the 5 'and 3' positions of the furanose ring of a nucleoside compound and form a prodrug form of the nucleoside compound. In another embodiment, the phosphoramidate present at the 5' position of the furanose ring of a nucleoside may form a cyclic phosphoramidate compound by forming a bond with the 3' -hydroxy substituent of the 3' position of the furanose ring of the nucleoside compound and form a prodrug form of the nucleoside compound.

The term "phosphorothioate", "phosphorothioate" or "phosphorothioate" is a moiety having a phosphorus bound to sulfur, two oxy groups and one amine (which may be optionally substituted). Phosphorothioates for use in the present invention are described in US patent No.8,772,474 and WO 2012/040124. Phosphorothioate groups useful in the present invention include those of the following structures:

other phosphorothioates include those of the following structure:

wherein:

R ^P1 is optionally taken out ofSubstituted linear, branched or cyclic alkyl, or optionally substituted aryl, heteroaryl or heterocyclic groups or a linked combination thereof; and

R ^P2 is-NR ^N1 R ^N2 A group or B';

wherein:

R ^N1 and R is ^N2 Each independently H, C ₁ -C ₈ Alkyl, (C) ₃ -C ₇ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, or (heteroaryl) C ₀ -C ₄ Alkyl-; or (b)

R ^N1 And R is ^N2 Taken together with the nitrogen atom to which it is attached, form a 3 to 7 membered heterocyclic ring;

b' isA group;

wherein:

R ¹⁶ is hydrogen, (C) ₁ -C ₈ ) Alkyl, (C) ₂ -C ₈ ) Alkenyl group (C) ₂ -C ₈ ) Alkynyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, (heteroaryl) C ₀ -C ₄ Alkyl-, or side chains of amino acids, for example side chains of amino acids generally selected from the group consisting of: alanine, beta-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine (typically, R ¹⁶ Hydrogen, methyl, isopropyl or isobutyl);

R ¹⁷ is hydrogen, (C) ₁ -C ₈ ) Alkyl, (C) ₂ -C ₈ ) Alkenyl group (C) ₂ -C ₈ ) Alkynyl, (C) ₃ -C ₈ Cycloalkyl) C ₀ -C ₄ Alkyl-, (aryl) C ₀ -C ₄ Alkyl-, (C) ₃ -C ₆ Heterocyclyl) C ₀ -C ₄ Alkyl-, (heteroaryl) C ₀ -C ₄ Alkyl-, or side chains of amino acids, for example side chains of amino acids generally selected from the group consisting of: alanine, beta-alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, glutamine, glycine, phenylalanine, histidine, isoleucine, lysine, leucine, methionine, proline, serine, threonine, valine, tryptophan or tyrosine (typically, R ¹⁷ Hydrogen, methyl, isopropyl or isobutyl);

R ¹⁸ is hydrogen or C ₁ -C ₃ An alkyl group; or (b)

B' isA group; and

R ¹⁸ 、R ¹⁹ 、R ²⁰ and R is ²¹ As defined above.

Preferred R ^P1 Groups include optionally substituted phenyl, naphthyl and monocyclic heteroaryl groups, particularly to increase the bioavailability of the compounds into the cells of a patient and to exhibit reduced toxicity Those groups (particularly lipophilic groups) that are sex, increased therapeutic index and increased pharmacokinetics (slower metabolism and excretion of the compound).

The thiocarbamate may be present at the 5 'or 3' position of the furanose ring of the nucleoside compound, forming a prodrug form of the nucleoside compound. In one embodiment, the thiocarbamates may be present at both the 5 'and 3' positions of the furanose ring of the nucleoside compound and form a prodrug form of the nucleoside compound. In another embodiment, phosphorothioates present at the 5' position of the furanose ring of a nucleoside may form cyclic phosphorothioate compounds by forming a bond with the 3' -hydroxy substituent at the 3' position of the furanose ring of the nucleoside compound and form a prodrug form of the nucleoside compound.

The term "D-configuration" as used in the context of the present invention refers to the main configuration of the natural structure of the mimic sugar moiety as opposed to the non-naturally occurring nucleoside or "L" configuration. The term "β" or "β anomer" is used to describe nucleoside analogs in which the nucleoside base is assembled (disposed) above the plane of the furanose moiety in the nucleoside analog.

The terms "co-administration" and "co-administration" or combination therapy are used to describe administration of at least one 2 '-deoxy-2' - α -fluoro-2 '- β -C-nucleoside compound according to the present invention in combination with at least one other active agent, such as at least one additional anti-HCV agent, where appropriate, including other 2' -deoxy-2 '- α -fluoro-2' - β -C-nucleoside agents disclosed herein. The timing of co-administration is preferably determined by the medical professional treating the patient. It is sometimes preferred to administer the agents simultaneously. Alternatively, the drugs selected for combination therapy may be administered to the patient at different times. Of course, when more than one virus or other infection or other condition is present, the compounds of the invention may be combined with other agents to treat other infections or conditions, as desired.

The term "host" as used herein refers to single or multicellular organisms, including cell lines and animals, typically humans, in which HCV virus can replicate. The term host refers in particular to infected cells, cells transfected with all or part of the HCV genome, as well as animals, particularly primates (including chimpanzees) and humans. In most animal applications of the invention, the host is a human patient. However, in certain instances, the invention obviously contemplates veterinary applications (e.g., chimpanzees). The host may be, for example, bovine, equine, avian, canine, feline, etc.

Isotopic substitution

The invention includes compounds and uses of compounds having the desired isotopic substitution of an atom, the amount of which is higher than the natural abundance of the isotope, i.e., enriched. Isotopes are atoms having the same atomic number but different mass numbers, i.e., the same number of protons but different numbers of neutrons. As a general example and not by way of limitation, isotopes of hydrogen, such as deuterium (2H) and tritium (3H), can be used anywhere in the structure. Alternatively or additionally isotopes of carbon may be used such as ¹³ C and C ¹⁴ C. Preferred isotopic substitution is substitution of deuterium for hydrogen at one or more positions on the molecule to improve the performance of the drug. Deuterium can bind near or beside the chain cleavage site (alpha-deuterium kinetic isotope effect) or the chain cleavage site during metabolism (beta-deuterium kinetic isotope effect). Achillion Pharmaceuticals, inc. (WO/2014/169278 and WO/2014/169280) describe that deuteration of nucleotides improves their pharmacokinetics or pharmacodynamics, including deuteration at the 5-position of the molecule.

Substitution with isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased half-life or reduced dosage requirements. Substitution of deuterium for hydrogen at the metabolic decomposition site can reduce the metabolic rate at the bond or eliminate metabolism at the bond. At any site of the compound where a hydrogen atom may be present, the hydrogen atom may be any isotope of hydrogen, including protium @, hydrogen atom ¹ H) Deuterium ² H) And tritium% ³ H) A. The invention relates to a method for producing a fibre-reinforced plastic composite Thus, unless the context clearly dictates otherwise, reference herein to a compound encompasses all possible isotopic forms.

The term "isotopically labeled" analog refers to its being "deuterated analog", " ¹³ C-labelled analogue "or" deuterated- ¹³ C-labeled analogues'Is a similar compound. The term "deuterated analog" refers to an analog in which the H-isotope, hydrogen/protium (1H), is replaced by the H-isotope, deuterium ² H) Substituted compounds described herein. Deuterium substitution may be partial or complete. Partial deuterium substitution refers to substitution of at least one hydrogen with at least one deuterium. In some embodiments, the isotope is isotopically enriched by 90, 95, or 99% at any location of interest. In certain embodiments, it is enriched in 90, 95 or 99% deuterium at the desired position. Deuterated is at least 80% at the selected position unless indicated to the contrary. Deuteration of nucleosides can occur at any substitutable hydrogen that provides the desired result. .

Methods of treatment or prophylaxis

Treatment as used herein is directed to the administration of an active compound to a host infected with HCV virus.

The term "prophylactic" or prophylaxis, when used, refers to the administration of an active compound to prevent or reduce the likelihood of occurrence of a viral disorder. The invention includes both therapeutic and prophylactic or preventative treatment. In one embodiment, the active compound is administered to a host that has been exposed to, and is therefore at risk of, infection by a hepatitis c virus.

The present invention relates to a method for treating or preventing the following diseases: hepatitis c virus (including drug resistant and multi-drug resistant forms of HCV and related disease states, conditions), or complications of HCV infection (including cirrhosis and related hepatotoxicity), as well as other conditions secondary to HCV infection (e.g., weakness, loss of appetite, weight loss, breast enlargement (especially in men), rashes (especially on palms), blood clotting difficulties, spider-like vessels on the skin, confusion, coma (encephalopathy), abdominal fluid accumulation (ascites), esophageal varices, portal hypertension, renal failure, splenomegaly, cytopenia, anemia, thrombocytopenia, jaundice, and hepatocellular carcinoma, among others). The method comprises administering to a host in need thereof an effective amount of at least one β -D-2'- α -fluoro-2' - β -C-substituted-2-modified-N as described herein ⁶ -substituted purine nucleotides, optionally in combination with at least one additional biologically active agent, e.g. an additional anti-HCV agent, further groupAnd pharmaceutically acceptable carrier additives and/or excipients.

In another aspect, the invention is a method of preventing or preventing: HCV infection or disease state or complications of related or secondary disease states, conditions, or HCV infection, including cirrhosis and related liver toxicity, frailty, anorexia, weight loss, breast enlargement (particularly in men), rash (particularly on palms of the hands), blood clotting difficulties, spider-like vessels on the skin, confusion, coma (encephalopathy), abdominal fluid accumulation (ascites), esophageal varices, portal hypertension, renal failure, splenomegaly, cytopenia, anemia, thrombocytopenia, jaundice, and hepatocellular carcinoma, and the like, comprising administering to a patient at risk an effective amount of at least one compound according to the invention as described above in combination with a pharmaceutically acceptable carrier, additive or excipient, optionally in combination with another anti-HCV agent. In another embodiment, the active compounds of the present invention may be administered to patients following hepatitis-related liver transplantation to protect new organs.

If desired, 5 '-stable beta-D-2' -D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N ⁶ The substituted purine nucleotide may be administered in the form of any salt or prodrug capable of providing the parent compound, directly or indirectly, or exhibiting activity per se when administered to a recipient. Non-limiting examples are pharmaceutically acceptable salts and compounds that have been modified at functional groups such as hydroxyl or amine functional groups to improve the biological activity, pharmacokinetics, half-life, controlled delivery, lipophilicity, absorption kinetics, ease of phosphorylation to the active 5' -triphosphate or delivery efficiency using the desired route of administration of the compound. Methods of improving the properties of an active compound to achieve a desired property are known to those skilled in the art or can be readily assessed by standard methods such as acylation, phosphorylation, phosphorothioamine phosphorylation, phosphoramidation, phosphorylation (phosphorylation), alkylation or pegylation.

IV pharmaceutical composition

In one aspect of the inventionIn one aspect, the pharmaceutical compositions according to the present invention comprise an anti-HCV virus effective amount of at least one 5' -stable β -D-2' - α -fluoro-2 ' - β -C-substituted-2-modified-N as described herein ⁶ -substituted purine nucleotide compounds, optionally in combination with pharmaceutically acceptable carriers, additives or excipients, further optionally in combination or alternation with at least one other active compound.

In one aspect of the invention, the pharmaceutical composition according to the invention comprises an anti-HCV virus effective amount of at least one active β -D-2'- α -fluoro-2' - β -C-substituted-2-modified-N as described herein ⁶ -substituted purine nucleotide compounds, optionally in combination with pharmaceutically acceptable carriers, additives or excipients, further optionally in combination with at least one other antiviral agent (e.g. an anti-HCV agent).

The present invention includes pharmaceutical compositions comprising a hepatitis C virus infection treating effective amount of a beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N of the present invention in a pharmaceutically acceptable carrier or excipient ⁶ -a substituted purine nucleotide compound or a salt or prodrug thereof. In another embodiment, the invention includes a pharmaceutical composition comprising an amount of a β -D-2'- α -fluoro-2' - β -C-substituted-2-modified-N of the invention in a pharmaceutically acceptable carrier or excipient effective to prevent hepatitis C virus infection ⁶ -a substituted purine nucleotide compound or a salt or prodrug thereof. .

One of ordinary skill in the art will recognize that the therapeutically effective amount will vary with the infection or disorder being treated, its severity, the treatment regimen being administered, the pharmacokinetics of the agent being used, and the patient or subject (animal or human) being treated, and that such therapeutic amount can be determined by the attending physician or practitioner.

5 '-stabilized beta-D-2' -D-2 '-alpha-fluoro-2' -beta-C-substituted-2-modified-N according to the invention ⁶ The substituted purine nucleotide compounds may be formulated in admixture with a pharmaceutically acceptable carrier. In general, it is preferred to administer the pharmaceutical composition in an oral administration form, but certain formulations may be administered parenterally, intravenously, intramuscularly, topically, transdermally, buccally, subcutaneously, by way of a suppositoryAgents or other routes of administration, including intranasal sprays. Intravenous and intramuscular formulations are typically administered in the form of sterile physiological saline. One of ordinary skill in the art can modify the formulation to render it more soluble in water or other vehicles, for example, this can be readily accomplished by minor modifications (salt preparation, esterification, etc.), which are well within the ordinary skill in the art. It is also well within the ordinary skill in the art to vary the route of administration and the particular compound regimen so as to control the pharmacokinetics of the compounds of the present invention to achieve the greatest beneficial effect in the patient. In certain pharmaceutical dosage forms, the prodrug forms of the compounds, including in particular acylated (acetylated or otherwise) and ether (alkyl and related) derivatives, phosphate esters, phosphorothioate phosphoramidates, phosphoramidates and various salts of the compounds of the present invention are preferred. One of ordinary skill in the art knows how to readily modify the compounds of the present invention into a prodrug form to facilitate delivery of the active compound to the host organism or to the target site of the patient. Those of ordinary skill in the art will also utilize the favorable pharmacokinetic parameters of the prodrug forms (when available) in delivering the compounds of the present invention to the host organism or to the target site of the patient to maximize the intended effect of the compound.

The amount of the compound included within the therapeutically active formulation according to the invention is an effective amount for treating HCV infection, reducing the likelihood of HCV infection, or inhibiting, reducing and/or eliminating HCV or secondary effects thereof, including disease states, disorders and/or complications secondary to HCV. In general, a therapeutically effective amount of a compound of the invention in a pharmaceutical dosage form will typically range from about 0.001mg/kg patient to about 100mg/kg patient or more per day, more typically, slightly less than about 0.1mg/kg to about 25mg/kg patient or significantly more, depending on the compound used, the condition or infection being treated, and the route of administration. The active nucleoside compounds according to the present invention are typically administered in an amount ranging from about 0.1mg/kg to about 15mg/kg of patient per day, depending on the pharmacokinetics of the agent in the patient. This dosage range generally yields an effective blood level concentration of the active compound, which may range from about 0.001 to about 100, from about 0.05 to about 100 micrograms/cc of blood in the patient.

Typically, to treat, prevent or delay the onset of such infections and/or reduce the likelihood of HCV viral infection or a secondary disease state, condition or complication of HCV, the composition should be administered in a total amount of about 250 micrograms up to about 500mg or more, at least once a day (e.g., at least 25, 50, 100, 150, 250 or 500 milligrams, up to four times a day) in oral dosage form. The compounds of the invention are generally administered orally, but may be administered parenterally, topically or in the form of suppositories, as well as intranasally, in the form of nasal sprays or as described elsewhere herein.

In the case of co-administration of a compound of the invention in combination with an additional anti-HCV compound as described elsewhere herein, the amount of the compound administered in accordance with the invention ranges from about 0.01mg/kg patient to about 500mg/kg patient or significantly more, depending on the co-administered second agent and its antiviral efficacy, the condition of the patient, and the severity of the disease or infection to be treated, and the route of administration. The other anti-HCV agents may be administered in an amount ranging, for example, from about 0.01mg/kg to about 500mg/kg. In certain preferred embodiments, these compounds may be administered in amounts typically ranging from about 0.5mg/kg to about 50mg/kg or more (typically up to about 100 mg/kg), typically depending on the pharmacokinetics of the two agents in the patient. These dosage ranges generally produce an effective blood level concentration of the active compound in the patient.

For the purposes of the present invention, a prophylactically or prophylactically effective amount of a composition according to the present invention falls within the same concentration ranges as the therapeutically effective amounts described above, and is generally the same as the therapeutically effective amount. .

Administration of the active compounds includes continuous (intravenous drip) to several times daily oral or intranasal administration (e.g., q.i.d.) or transdermal administration, and may include oral, topical, parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include penetration enhancers), buccal and suppository administration, and other routes of administration. To enhance the bioavailability of the oral route compounds, enteric coated oral tablets may also be used. The most effective dosage form will depend on the bioavailability/pharmacokinetics of the particular agent selected and the severity of the disease in the patient. Oral dosage forms are particularly preferred because of ease of administration and anticipated favorable patient compliance.

To prepare the pharmaceutical compositions according to the present invention, a therapeutically effective amount of one or more compounds according to the present invention is typically thoroughly mixed with a pharmaceutically acceptable carrier to produce a dose according to conventional pharmaceutical mixing techniques. The carrier may take a variety of forms depending on the form of formulation desired for administration, such as oral or parenteral administration. In preparing the pharmaceutical composition in oral dosage form, any of the usual pharmaceutical media may be employed. Thus, for liquid oral formulations, such as suspensions, elixirs and solutions, suitable carriers and additives including water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like may be employed. For solid oral formulations, such as powders, tablets, capsules, and for solid formulations such as suppositories, suitable carriers and additives including starches, sugar carriers such as dextrose, mannitol (manifold), lactose and related carriers, diluents, granulating agents, lubricants, binders, disintegrating agents and the like can be used. If desired, the tablets or capsules may be enteric coated or sustained release by standard techniques. The use of these dosage forms can significantly increase the bioavailability of the compound in a patient.

For parenteral formulations, the carrier will typically comprise sterile water or aqueous sodium chloride solution, but may also include other ingredients, including those which aid in dispersion. Of course, when sterile water is used and remains sterile, the compositions and carriers must also be sterilized. Injectable suspensions may also be prepared in which case suitable liquid carriers, suspending agents and the like may be employed.

Liposomal suspensions (including liposomes targeted to viral antigens) can also be prepared by conventional methods for producing pharmaceutically acceptable carriers. This may be suitable for delivering the free nucleoside, acyl/alkyl nucleoside or phosphate ester prodrug form of the nucleoside compound according to the present invention.

In typical embodiments according to the invention, the compounds and compositions are used to treat, prevent or delay HCV infection or a secondary disease state, disorder or complications of HCV.

V. combination and alternation therapy

It has been recognized that drug resistant mutants of viruses can appear after prolonged treatment with antiviral agents. Drug resistance most commonly occurs due to mutations in genes encoding enzymes used in viral replication. The efficacy of a drug against HCV infection can be prolonged, enhanced or restored by the combined or alternating administration of the compound with another, and perhaps even two or three other antiviral compounds that induce a mutation that is different from the primary drug-induced mutation or act via a different pathway than the primary compound pathway. Alternatively, the pharmacokinetics, biodistribution, half-life or other parameters of the drug may be altered by such combination therapy (which may include alternating therapies if deemed consistent). Due to the disclosed beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N ⁶ The substituted purine nucleotides are NS5B polymerase inhibitors, then it is useful to administer the compound in combination with, for example:

(1) Protease inhibitors, such as NS3/4A protease inhibitors;

(2) NS5A inhibitors;

(3) Another NS5B polymerase inhibitor;

(4) NS5B non-substrate inhibitors;

(5) Interferon alpha-2 a, which may be pegylated or otherwise modified, and/or ribavirin;

(6) A non-substrate-based inhibitor;

(7) An helicase inhibitor;

(8) Antisense oligodeoxynucleotides (S-ODNs);

(9) An aptamer;

(10) Nuclease-drug-resistant ribozyme;

(11) iRNA, including microRNA and SiRNA;

(12) Antibodies, partial antibodies or domain antibodies to viruses, or

(13) Viral antigens or partial antigens that induce a host antibody response.

Can be substituted with the beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-of the present invention-2-modified-N ⁶ Non-limiting examples of anti-HCV agents administered in combination with substituted purine nucleotides are:

(i) Protease inhibitors, e.g. telaprevirBessepir (Victrelis) ^TM ) Cimicavir (Olysio) ^TM ) Paritaprevir (ABT-450), ACH-2684, AZD-7295; BMS-791325; danoprevir; filibrevir; GS-9256; GS-9451; MK-5172; setrobrevir; sovaprevir; tegobovir; VX-135; VX-222 and ALS-220;

(ii) NS5A inhibitors such as ACH-2928, ACH-3102, IDX-719, dacatasvir, ledispasvir and Ombitasvir (ABT-267);

(iii) NS5B inhibitors such as ACH-3422; AZD-7295; clemizole; ITX-5061; PPI-461; PPI-688,MK-3682 and mericitabine;

(iv) NS5B inhibitors, such as ABT-333, MBX-700; and

(v) Antibodies, such as GS-6624.

If beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N ⁶ Substituted purine nucleotides for use in the treatment of advanced hepatitis c virus leading to liver cancer or cirrhosis, which may be combined or alternatively administered in one embodiment with another drug commonly used in the treatment of hepatocellular carcinoma (HCC), for example as described in Andrew Zhu in "New Agents on the Horizon in Hepatocellular Carcinoma" Therapeutic Advances in Medical Oncology, V5 (1), january 2013,41-50. Examples of suitable compounds for combination therapy when the host has or is at risk of HCC include anti-angiogenic agents, sunitinib, brianinib, linifanib, ramucirumab, bevacizumab, cerinib, pazopanib, TSU-68, lenvatinib, anti-EGFR antibodies, mTor inhibitors, MEK inhibitors and histones Deacetylation inhibitors.

The drugs currently approved for influenza are amantadine, rimantadine and oseltamivir. Any of these agents may be used in combination or alternation with the active compounds provided herein for the treatment of viral infections susceptible thereto. Ribavirin is used to treat measles, influenza a, influenza B, parainfluenza, severe RSV bronchiolitis and SARS, and other viral infections, and thus is particularly useful in combination with the compounds of the invention for treating hosts infected with single stranded RNA viruses. Palivizumab is approved for infants at high risk of RSV infection.

Currently, there is no approved drug for west nile virus. Physicians recommend providing intensive supportive treatment that may include hospitalization, intravenous infusion, respiratory assistance with ventilators, drug control of seizures, brain swelling, nausea and vomiting, and antibiotic prevention of bacterial infection that makes the disease worse. This strengthens the importance of the compounds of the invention for use in viral medical therapy.

VI beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N of the invention ⁶ Process for the preparation of substituted purine nucleotides

General methods for providing the compounds of the present invention are known in the art or described herein. The synthesis of 2' -chloro nucleotides is described in US 20150366888, WO 2014058801; WO 2015/066370 and WO 2015200219.

The following abbreviations are used in the synthesis schemes.

CBr ₄ Carbon tetrabromide

DBU 1, 8-diazabicyclo [5.4.0] undec-7-ene

DCM: dichloromethane

THF: tetrahydrofuran (THF), anhydrous

EtOAc: ethyl acetate

EtOH/ethanol

Li(OtBu) ₃ AlH lithium aluminum tri-tert-butoxide

Na ₂ SO ₄ Sodium sulfate (Anhydrous)

MeCN: acetonitrile

MeNH ₂ Methyl amine

MeOH: methanol

Na ₂ SO ₄ Sodium sulfate

NaHCO ₃ Sodium bicarbonate

NH ₄ Cl: ammonium chloride

NH ₄ OH: ammonium hydroxide

PE: petroleum ether

Ph ₃ P: triphenylphosphine

Silica gel (230 to 400 mesh, adsorbent)

t-BuMgCl: t-butylmagnesium chloride

t-BuOK: sodium tert-butoxide

t-BuOH: t-Butanol

Examples

General method

¹ H、 ¹⁹ F and F ³¹ The P NMR spectrum was recorded on a 300MHz Fourier transform Brucker spectrometer. The spectrum is from that of CDCl ₃ 、CD ₃ OD or DMSO-d ₆ Obtained as a sample prepared from a 5mm diameter tube. Spin multispeed is indicated by the following symbols: s (singlet), d (doublet), t (triplet), m (multiplet) and Br (broad). Coupling constants (J) are reported in Hz. MS spectra were obtained using electrospray ionization (ESI) at Agilent Technologies 6120 quadrupole MS apparatus. The reaction is typically carried out under a dry nitrogen atmosphere using Sigma-Aldrich anhydrous solvent. All commonly used chemicals were purchased from commercial sources. .

i)Li(OtBu) ₃ AlH，THF，-30℃--＞-15℃；ii)PPh ₃ ，CBr ₄ DCM, -20 ℃ to > 0 ℃; iii) 2-amino-6-chloropurine, tBuOK, tBuOH/MeCN 9:1, 65 ℃; iv) MeNH ₂ (33%), meOH,85 ℃; v) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF,0 ℃ C. - > room temperature

EXAMPLE 1 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester

Preparation of((2R, 3R,4R, 5R) -3- (benzoyloxy) -5-bromo-4-fluoro-4-methyltetrahydrof-n-2-yl) methylbenzoate (2)

To a solution of (2R) -3, 5-di-O-benzoyl-2-fluoro-2-C-methyl-D-ribono-gamma-lactone (24.8 g,66.6 mmol) in dry THF (333 mL) under nitrogen and cooled to-30℃was added lithium aluminum tri-tert-butoxide (1.0M in THF, 22.6mL,22.6 mmol) dropwise. After the addition was complete, the reaction mixture was slowly warmed to-15 ℃ over 90 minutes, then EtOAc (300 mL) was added and saturated NH ₄ The mixture was quenched with aqueous Cl (200 mL). The obtained solution is treated in the following conditionThe filtrate was filtered and extracted twice with EtOAc. The combined organics were dried (Na ₂ SO ₄ ) Filtered and concentrated. The residue was collected in anhydrous DCM (225 mL) under nitrogen, cooled to-20deg.C, and PPh was then added ₃ (19.1 g,72.8 mmol). After stirring at-20℃for 10 minutes, CBr was added ₄ (26.0 g,78.4 mmol) and the reaction mixture was slowly warmed to 0℃over 2 hours. The resulting mixture was poured onto a silica gel column and eluted with PE/EtOAc (gradient 100:0 to 80:20). Fractions containing α -bromofuranoside were collected and concentrated to give product 2 (18.1 g,41.3mmol, 62% over 2 steps) as a thick colorless oil.

¹ H NMR(300MHz,CDCl ₃ )δ8.15-8.11(m,2H),8.04-8.01(m,2H),7.64-7.55(m,2H),7.51-7.41(m,4H),6.34(d,J＝1.6Hz,1H),5.29(dd,J＝5.5,3.1Hz,1H),4.89-4.85(m,1H),4.78(dd,J＝12.5,3.2Hz,1H),4.63(dd,J＝12.5,4.5Hz,1H),1.72(d,J＝21.6Hz,3H). ¹⁹ F NMR(282MHz,CDCl ₃ )δ-150.0.

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (benzoyloxymethyl) -4-fluoro-4-methyltetrahydrof-n-3-ylbenzoate (3)

2-amino-6-chloropurine (2.63 g,15.5 mmol) was suspended in t-BuOH (54 mL) under nitrogen. The reaction mixture was heated to 30℃and potassium tert-butoxide (1.69 g,15.1 mmol) was then added. After 45 minutes, a solution of bromofuranoside 2 (2.24 g,5.12 mmol) in anhydrous MeCN (6 mL) was added and the reaction mixture was heated to 65 ℃ for 16 hours and then cooled to room temperature. Adding saturated NH ₄ Aqueous Cl (70 mL) and the resulting solution was extracted with EtOAc (3X 60 mL). The combined organics were dried (Na ₂ SO ₄ ) Filtered and concentrated. The residue was purified twice by column chromatography (gradient PE/EtOAc 80:20 to 0:100, then 60:40 to 20:80) to give product 3 (1.56 g,2.96mmol, 57%) as a white solid.

¹ H NMR(300MHz,CDCl ₃ )δ8.05-8.02(m,2H),7.95-7.92(m,2H),7.88(s,1H),7.63-7.57(m,1H),7.53-7.41(m,3H),7.35-7.30(m,2H),6.43(dd,J＝22.6,9.1Hz,1H),6.12(d,J＝18.3Hz,1H),5.34(br s,2H),5.00(dd,J＝11.9,4.5Hz,1H),4.79-4.73(m,1H),4.60(dd,J＝11.9,5.3Hz,1H),1.34(d,J＝22.6Hz,3H). ¹⁹ F NMR(282MHz,CDCl ₃ )δ-157.0.MS(ESI)m/z C ₂₅ H ₂₂ FN ₅ O ₅ [M+H] ⁺ Theoretical value: 526.9; actual measurement value: 527.0.

preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-n-3-ol (4)

To a solution of compound 3 (575 mg,1.09 mmol) in MeOH (9 mL) was added methylamine (33% in anhydrous EtOH, 1.7mL,1.81 mmol). The reaction mixture was heated to 85 ℃ in a sealed tube for 16 hours, cooled to room temperature and concentrated. First by column chromatography (gradient DCM/MeOH 100:0 to 85:15) followed by reverse phase column chromatography (gradient H) ₂ The residue was purified with O/MeOH 100:0 to 0:100 to give product 4 (284 mg,0.91mmol, 84%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD)δ8.06(s,1H),6.11(d,J＝18.1Hz,1H),4.41(dd,J＝24.4,9.1Hz,1H),4.07-4.01(m,2H),3.86(dd,J＝12.9,3.3Hz,1H),3.04(br s,3H),1.16(d,J＝22.3Hz,3H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.7.MS(ESI)m/z C ₁₂ H ₁₉ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 313.1; actual measurement value: 313.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (5)

To a solution of compound 4 (114 mg, 365. Mu. Mol) in anhydrous THF (4 mL) under nitrogen and cooled to 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 0.66mL, 660. Mu. Mol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes, then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and then a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (Ross, b.s., reddy, p.g., zhang, h.r., rachakonda, s., and Sofia, m.j., j.org., chem., (2011), (255 mg,558 μmol) in anhydrous THF (1 mL) was added dropwise over 10 minutes the reaction mixture was stirred at 0 ℃ for 30 minutes, then stirred at room temperature for 18 hours, then saturated NH ₄ Aqueous Cl (4 mL) was quenched and extracted with EtOAc (3X 5 mL). The combined organics were dried, filtered (Na ₂ SO ₄ ) And concentrated. First by column chromatography (gradient DCM/MeOH 100:0 to 90:10) followed by reverse phase column chromatography (gradient H) ₂ The residue was purified with O/MeOH 100:0 to 0:100) to give product 5 (mixture of diastereomers, 101mg, 174. Mu. Mol, 48%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD) delta 7.83 (s, 0.55H), 7.82 (s, 0.45H), 7.38-7.16 (m, 5H), 6.15 (d, j=18.5 hz, 0.45H), (d, j=18.8 hz, 0.55H), 4.99-4.88 (with H) ₂ O overlap, m, 1H), 4.65-4.36 (m, 3H), 4.25-4.17 (m, 1H), 3.97-3.85 (m, 1H), 3.05 (br s, 3H), 1.32-1.28 (m, 3H), 1.25-1.15 (m, 9H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-162.8(s),-163.3(s). ³¹ P NMR(121MHz,CD ₃ OD)δ4.10(s),3.99(s).MS(ESI)m/z C ₂₄ H ₃₄ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 582.2; actual measurement value: 582.2.

i)Me ₂ NH.HCl, DBU, meOH,85 ℃; v) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L, isopropyl alaninate, tBungCl, THF, 0deg.C.

EXAMPLE 2 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (7)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-n-3-ol (6)

To a solution of compound 3 from example 1 (500 mg,0.95 mmol) in MeOH (6 mL) was added dimethylamine hydrochloride (783 mg,9.6 mmol) and 1, 8-diazabicyclo [ 5.4.0)]Undec-7-ene (1.43 mL,9.6 mmol). The reaction mixture was heated in a sealed tube at 85 ℃ for 6 hours, cooled to room temperature and concentrated. First by column chromatography (gradient DCM/MeOH 100:0 to 85:15) followed by reverse phase column chromatography (gradient H) ₂ The residue was purified with O/MeOH 100:0 to 0:100 to give product 6 (200 mg,0.61mmol, 64%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD)δ8.07(s,1H),6.14(d,J＝18.1Hz,1H),4.41(dd,J＝24.4,9.2Hz,1H),4.08-4.02(m,2H),3.87(dd,J＝12.8,2.9Hz,1H),3.42(br s,6H),1.16(d,J＝22.0Hz,3H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.8.MS(ESI)m/z C ₁₃ H ₂₀ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 327.2; actual measurement value: 327.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (7)

To a solution of compound 6 (80 mg, 245. Mu. Mol) in anhydrous THF (4 mL) under nitrogen and cooled to 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 0.64mL, 640. Mu. Mol) over 10 min. Stirring at 0deg.CThe reaction mixture was stirred at room temperature for 15 minutes and then for another 15 minutes. The reaction mixture was cooled to 0℃and then a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (167 mg, 367. Mu. Mol) in anhydrous THF (4 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were dried, filtered (Na ₂ SO ₄ ) And concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) followed by reverse phase column chromatography (gradient H2O/MeOH 100:0 to 0:100) to give product 7 (mixture of diastereomers, 35mg, 58. Mu. Mol, 24%) as a white solid.

¹ H NMR(300MHz，CD ₃ OD) delta 7.83 (s, 0.5H), 7.82 (s, 0.5H), 7.34-7.16 (m, 5H), 6.15 (d, j=18.7 hz, 0.5H), 6.13 (d, j=18.8 hz, 0.5H), 4.99-4.85 (with H) ₂ O overlap, m, 1H), 4.65-4.26 (m, 3H), 4.27-4.12 (m, 1H), 3.99-3.81 (m, 1H), 3.42,3.41 (2 b r s, 6H), 1.36-1.25 (m, 3H), 1.24-1.11 (m, 9H). ¹⁹ F NMR(282MHz，CD ₃ OD)δ-162.7(s)，-163.2(s). ³¹ P NMR(121MHz，CD ₃ OD)δ4.08(s)，4.00(s).MS(ESI)m/z C ₂₅ H ₃₆ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 596.5; actual measurement value: 596.2.

i) a) N-methylcyclopropylamine hydrochloride, et ₃ N，MeOH，100℃；b)NH ₄ OH, meOH,100 ℃; ii) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF, 0deg.C.

EXAMPLE 3 preparation of((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-ol-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (9)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclopropylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-N-3-ol (8)

To a solution of compound 3 (600 mg,1.14 mmol) in MeOH (10 mL) was added N-methylcyclopropylamine hydrochloride (365 mg,3.40 mmol) and triethylamine (470 μl,3.40 mmol). The reaction mixture was heated at 100 ℃ in a sealed tube for 15 hours and cooled to room temperature. Adding a catalyst containing 30% NH ₄ Aqueous OH (4 mL) and the reaction mixture was heated in a sealed tube at 100 ℃ for 2 hours, cooled and concentrated. The residue was purified by column chromatography (gradient: DCM/MeOH100:0 to 90:10) to give product 8 (351 mg,0.99mmol, 87%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD)δ8.13(s,1H),6.15(d,J＝18.0Hz,1H),4.40(dd,J＝24.3,9.0Hz,1H),4.06-4.02(m,2H),3.89-3.83(m,1H),3.32(m,3H),3.18-3.11(m,1H),1.16(d,J＝22.2Hz,3H),0.96-0.89(m,2H),0.74-0.69(m,2H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.8.MS(ESI)m/z C ₁₅ H ₂₂ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 353.2; actual measurement value: 353.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-ol-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (9)

To a solution of compound 8 (200 mg,0.57 mmol) in anhydrous THF (15 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 680 μl,0.68 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes and then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (283 mg,0.62 mmol) in anhydrous THF (4 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). Through Na ₂ SO ₄ The combined organics were dried and concentrated. First by column chromatography (gradientDCM/MeOH 100:0 to 90:10) followed by reverse phase column chromatography (gradient H ₂ The residue was purified with O/MeOH 100:0 to 0:100 to give product 9 (mixture of diastereomers, 160mg,0.26mmol, 45%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD) delta 7.85 (m, 1H), 7.38-7.16 (m, 5H), 6.18 (d, j=18.6 Hz) and 6.16 (d, j=18.9 Hz, 1H), 4.95-4.90 (with H) ₂ O overlap, m, 1H), 4.58-4.47 (m, 3H), 4.22-4.19 (m, 1H), 3.95-3.87 (m, 1H), 3.36-3.34 (overlap with MeOH, m, 3H), 3.19-3.12 (m, 1H), 1.32-1.22 (m, 12H), 0.96-0.89 (m, 2H), 0.74-0.69 (m, 2H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.11(s),4.02(s).MS(ESI)m/z C ₂₇ H ₃₈ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 622.2; actual measurement value: 622.2.

i) 2, 6-dichloropurine, tBuOK, tBuOH/MeCN,65 ℃; ii) MeNH ₂ MeOH,130 ℃; iii) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF,0℃to RT

EXAMPLE 4 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2, 6-dimethylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (12)

Preparation of (2 r,3r,4r,5 r) -5- (2, 6-dichloro-9H-purin-9-yl) -2- (benzoyloxymethyl) -4-fluoro-4-methyltetrahydrof-n-3-ylbenzoate (10).

The compound 2, 6-dichloropurine (1.30 g,6.86 mmol) was suspended in t-BuOH (25 mL) under nitrogen. Potassium tert-butoxide (778 mg,6.92 mmol) was added in portions, and the reaction mixture was stirred at room temperature. After 1 hour, a solution of bromofuranoside 2 (1.0 g,2.29 mmol) in anhydrous MeCN (20 mL) was added and the reaction mixture was heated at 65 ℃ overnight and then cooled to room temperature. Adding saturated NH ₄ Aqueous Cl and the resulting solution was extracted with EtOAc (3 times). Combining the organic mattersThrough Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient PE/EtOAc 100:0 to 0:100) to give product 10 (148 mg,0.27mmol, 12%) as a viscous solid.

¹ H NMR(300MHz，CDCl ₃ )δ8.31(s，1H)，8.12-8.09(m，2H)，8.02-7.99(m，2H)，7.64-7.39(m，6H)，6.38(d，J＝17.2Hz，1H)，6.02(dd，J＝21.2，8.9Hz，1H)，4.90-4.68(m，3H)，1.33(d，J＝22.4Hz，3H). ¹⁹ F NMR(282MHz，CDCl ₃ )δ-158.0.MS(ESI)m/z C ₂₅ H ₂₀ Cl ₂ FN ₄ O ₅ [M+H] ⁺ Is the theoretical value of (3): 546.4; actual measurement value: 546.3.

preparation of (2R, 3R,4R, 5R) -5- (2, 6-dimethylamino-9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrofuran-3-ol (11)

A solution of compound 10 (148 mg,0.27 mmol) in methylamine (33% in EtOH, 30 mL) was heated in a sealed tube at 130℃for 4 days, cooled to room temperature and concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 50:50) followed by reverse phase column chromatography (gradient H2O/MeOH 100:0 to 0:100) to give product 11 (33 mg,0.10mmol, 37%) as a white solid. ¹ H NMR(300MHz,CD ₃ OD)δ8.00(s,1H),6.12(d,J＝18.5Hz,1H),4.51(dd,J＝24.4,9.5Hz,1H),4.06-3.85(m,3H),3.04(s,3H),2.93(s,3H),1.20(d,J＝22.4Hz,3H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.2.MS(ESI)m/z C ₁₃ H ₂₀ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 327.2; actual measurement value: 327.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2, 6-dimethylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (12)

To a solution of compound 11 (55 mg,0.17 mmol) in anhydrous THF (2 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1M in THF, 304mL,0.30 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes and then at room temperature for 15 minutes. The solution was cooled to 0℃and added dropwise over 10 minutesA solution of (((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (115 mg,0.25 mmol) in dry THF (1 mL). The mixture was allowed to warm slowly to room temperature and stirred for 4 days. With saturated NH ₄ The reaction was quenched with aqueous Cl and extracted with EtOAc (3 times). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 50:50) to give product 12 (mixture of diastereomers, 13mg,0.02mmol, 13%) as a white solid. ¹ H NMR(300MHz,CD ₃ OD) delta 7.78 (s, 1H), 7.35-7.12 (m, 5H), 6.13 (d, j=19.1 hz, 0.53H), 6.10 (d, j=19.2 hz, 0.47H), 4.99-4.78 (overlapping H2O, m, 1H), 4.72-4.46 (m, 3H), 4.24-4.15 (m, 1H), 3.79-3.92 (m, 1H), 3.02 (br s, 3H), 2.92 (s+s, 3H), 1.29-1.11 (m, 12H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-162.0(s),-162.3(s). ³¹ P NMR(121MHz,CD ₃ OD)δ3.97(s),3.89(s).MS(ESI)m/z C ₂₅ H ₃₆ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 596.6; actual measurement value: 596.2.

i)TIPDSCl ₂ imidazole, DMF; ii) isobutyryl chloride, pyridine; iii) TBAF, THF; iv) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF, 0deg.C.

EXAMPLE 5 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-isobutyrylamino-6-methylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (16)

Step 1 preparation of Compound 13

To a solution of compound 4 (284 mg,0.92 mmol) and imidazole (370 mg,5.43 mmol) in anhydrous DMF (6 mL) at 0deg.C was added 1, 3-dichloro-1, 3-tetraisopropyl disiloxane (300 μl,0.94 mmol). The reaction mixture was stirred at room temperature for 2 hours, diluted with EtOAc (50 mL) and saturated NH ₄ Aqueous Cl and brine (40 mL each) were used to wash the suspensionAnd (3) liquid. Subjecting the organic material to Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient PE/EtOAc 7:3 to 3:7) to give product 13 (283 mg,0.51mmol, 56%) as a white solid. MS (ESI) m/z C ₂₄ H ₄₄ FN ₆ O ₄ Si ₂ [M+H] ⁺ Is the theoretical value of (3): 555.8; actual measurement value: 555.2.

step 2 preparation of Compound 14

To a solution of compound 13 (200 mg,0.36 mmol) in anhydrous pyridine (3 mL) at 0deg.C was added isobutyryl chloride (38 μl,0.36 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched by the addition of water (500 μl). The mixture was concentrated and co-evaporated with toluene (3X 10 mL). The residue was purified by column chromatography (gradient PE/EtOAc 1:0 to 1:1) to give product 14 (99 mg,0.16mmol, 44%) as a white solid. MS (ESI) m/z C ₂₈ H ₅₀ FN ₆ O ₅ Si ₂ [M+H] ⁺ Is the theoretical value of (3): 625.9; actual measurement value: 625.3.

preparation of (2R, 3R,4R, 5R) -5- (2-isobutyrylamino 6-methylamino-9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-n-3-ol (15)

To a solution of compound 14 (90 mg,0.14 mmol) in anhydrous THF (2 mL) was added tetrabutylammonium fluoride (1 m in THF, 38 μl,0.38 mmol). The mixture was stirred at room temperature for 2 hours and concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 10:0 to 9:1) followed by reverse phase column chromatography (gradient H2O/MeOH 100:0 to 0:100) to give product 15 (42 mg,0.11mmol, 77%) as a white solid. ¹ H NMR(300MHz,CD ₃ OD)δ8.31(s,1H),6.29(d,J＝17.9Hz,1H),4.70-4.60(m,1H),4.07-3.98(m,2H),3.89(dd,J＝12.5,3.4Hz,1H),3.10(br s,3H),2.87(br s,1H),1.23-1.16(m,9H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.8.MS(ESI)m/z C ₁₆ H ₂₄ FN ₆ O ₄ [M+H] ⁺ Is the theoretical value of (3): 383.4; actual measurement value: 383.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-isobutyrylamino 6-methylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (16)

To a solution of compound 15 (27 mg,0.07 mmol) in anhydrous THF (1 mL) at 0deg.C was added dropwise butyl magnesium chloride (1.0M in THF, 130 μL,0.13 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes, then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (50 mg,0.11 mmol) in anhydrous THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0℃for 30 min, then at room temperature for 18 h, then with saturated NH ₄ The Cl solution (2 mL) was quenched and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 95:5) followed by reverse phase column chromatography (gradient H2O/MeOH 100:0 to 0:100) to give product 16 (mixture of 2 diastereomers, 25mg,0.04mmol, 54%) as a white solid. ¹ H NMR(300MHz,CD ₃ OD) delta 8.05 (s, 1H), 7.33-7.13 (m, 5H), 6.27 (d, j=18.6 Hz) and 6.21 (d, j=19.1 Hz, 1H), 5.10-4.95 (m, 1H), 4.93-4.78 (overlapping H2O, m, 1H), 4.60-4.42 (m, 2H), 4.26-4.18 (m, 1H), 3.90-3.80 (m, 1H), 3.09 (br s, 3H), 2.84-2.80 (m, 1H), 1.33-1.15 (m, 18H). ³¹ P NMR(121MHz,CD ₃ OD)δ3.69(s). ³¹ P NMR(121MHz,CD ₃ OD)δ4.11(s),3.99(s).MS(ESI)m/z C ₂₈ H ₄₀ FN ₇ O ₈ P[M+H] ⁺ Is the theoretical value of (3): 652.6; actual measurement value: 652.3.

i) N-methylethylamine, meOH, 100deg.C; ii) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBungCl, THF, 0deg.C.

EXAMPLE 6 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-ethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-ol-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (18)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-ethylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-N-3-ol (17)

To a solution of compound 3 (150 mg,0.29 mmol) in MeOH (4 mL) was added N-methylethylamine (245. Mu.L, 2.90 mmol). The reaction mixture was heated at 100 ℃ in a sealed tube for 15 hours, cooled to room temperature and concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give product 31 (89 mg,0.26mmol, 89%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD)δ8.06(s,1H),6.13(d,J＝18.0Hz,1H),4.40(dd,J＝24.9,8.7Hz,1H),4.11-4.01(m,4H),3.98-3.83(m,1H),3.34(br.s,3H),1.24-1.11(m,6H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.7.MS(ESI)m/z C ₁₄ H ₂₂ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 341.2; actual measurement value: 341.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-ethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-ol-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (18)

To a solution of compound 17 (30 mg,0.09 mmol) in anhydrous THF (2 mL) at 0 ℃ was added dropwise t-butylmagnesium chloride (1.0 m in THF, 110 μl,0.11 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes, then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (48 mg,0.11 mmol) in anhydrous THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give product 18 (mixture of 2 diastereomers, 22mg,0.04mmol, 40%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD) delta 7.69 (m, 1H), 7.26-7.04 (m, 5H), 6.05 (d, j=18.6 Hz) and 6.03 (d, j=18.9 Hz, 1H), 4.86-4.79 (with H) ₂ O overlap, m, 1H), 4.50-4.32 (m, 3H), 4.12-4.06 (m, 1H), 3.96-3.79 (m, 3H), 3.25 (br.s, 3H), 1.24-1.02 (m, 15H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.07(s),4.00(s).MS(ESI)m/z C ₂₆ H ₃₈ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 609.3; actual measurement value: 609.2.

i) N-methylpropylamine, meOH, 100deg.C; ii) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF, 0deg.C.

EXAMPLE 7 preparation of((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-propylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (20)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-propylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrofuran-3-ol (19)

To a solution of compound 3 (150 mg,0.29 mmol) in MeOH (4 mL) was added N-methylpropylamine (295. Mu.L, 2.90 mmol). The reaction mixture was heated in a sealed tube at 100 ℃ for 15 hours, cooled to room temperature and concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) followed by reverse phase column chromatography (gradient H2O/MeOH 100:0 to 0:100) to give the product 19 as a white solid (80 mg,0.23mmol, 78%).

¹ H NMR(300MHz,CD ₃ OD) δ8.04 (s, 1H), 6.13 (d, j= 18.3,1H), 4.40 (dd, j=24.2, 9.2hz, 1H), m,4.06-3.84 (m, 5H), 1.68 (heptapeak, j=7.5 hz, 2H), 1.15 (d, j=22.2 hz, 3H), 0.93 (t, j=7.5 hz, 3H). ¹⁹ F NMR(282MHz,CD ₃ OD)δ-163.8.MS(ESI)m/z C ₁₅ H ₂₄ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 355.2; actual measurement value: 355.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-propylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (20)

To a solution of compound 19 (30 mg,0.09 mmol) in anhydrous THF (2 mL) at 0 ℃ over 10 min, t-butylmagnesium chloride (1.0 m in THF, 110 μl,0.11 mmol) was added dropwise. The reaction mixture was stirred at 0 ℃ for 15 minutes, then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (46 mg,0.11 mmol) in anhydrous THF (1 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with Cl solution (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give product 20 (mixture of 2 diastereomers, 22mg,0.03mmol, 33%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD) delta 7.78,7.77 (s+s, 1H), 7.37-7.13 (m, 5H), 6.15 (d, j=18.6 Hz) and 6.13 (d, j=18.9 Hz, 1H), 4.97-4.89 (with H) ₂ O overlap, m, 1H), 4.63-4.30 (m, 3H), 4.22-4.14 (m, 1H), 4.02-3.84 (m, 2H), 1.74-1.63 (3H, m), 1.32-1.27 (m, 3H), 1.23-1.13 (m, 9H), 0.94 (t, j=7.4 Hz) and 0.93 (t, j=7.4 Hz, 3H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.05(s),4.00(s).MS(ESI)m/z C ₂₇ H ₄₀ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 623.3; actual measurement value: 623.2.

i) a) N-methylcyclobutylamine hydrochloride, et ₃ N，MeOH，100℃；b)NH ₄ OH, meOH,100 ℃; ii) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF, 0deg.C.

EXAMPLE 8 preparation of((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclobutylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (22)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclobutylamino) -9H-purin-9-yl) -4-fluoro-2- (hydroxymethyl) -4-methyltetrahydrof-N-3-ol (21)

To a solution of compound 3 (150 mg,0.29 mmol) in MeOH (4 mL) were added N-methylcyclobutylamine hydrochloride (105 mg,0.90 mmol) and triethylamine (190. Mu.L, 1.00 mmol). The reaction mixture was heated at 100 ℃ in a sealed tube for 15 hours and cooled to room temperature. Adding a catalyst containing 30% NH ₄ An aqueous solution of OH (1 mL) and the reaction mixture was heated at 100 ℃ in a sealed tube for 2 hours, cooled and concentrated. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give product 21 (90 mg,0.25mmol, 86%) as a pale yellow solid.

¹ H NMR(300MHz，CD ₃ OD)δ8.09(s，1H)，6.14(d，J＝18.0Hz，1H)，5.80-5.70(m，1H)，4.44-4.33(m，1H)，4.06-4.02(m，2H)，3.88-3.84(m，1H)，3.34(s，3H)，2.38-2.19(m，4H)，1.79-1.71(m，2H)，1.17(d，J＝22.2Hz，3H). ¹⁹ F NMR(282MHz，CD ₃ OD)δ-163.8.MS(ESI)m/z C ₁₆ H ₂₄ FN ₆ O ₃ [M+H] ⁺ Is the theoretical value of (3): 367.2; actual measurement value: 367.2.

preparation of((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methyl-cyclobutylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-N-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (22)

To a solution of compound 21 (50 mg,0.14 mmol) in anhydrous THF (2 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 210 μl,0.21 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes and then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (74 mg,0.16 mmol) in anhydrous THF (2 mL) was added dropwise over 10 minutes. Stirring the reaction mixture at 0 ℃30 minutes and stirred at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) and then purified by reverse phase column chromatography (gradient H ₂ The residue was purified with O/MeOH 100:0 to 0:100) to give product 22 (mixture of 2 diastereomers, 24mg,0.04mmol, 28%) as a white solid.

¹ H NMR(300MHz,CD ₃ OD) delta 7.79 (s, 0.2H), 7.77 (s, 0.8H), 7.38-7.12 (m, 5H), 6.18 (d, j=17.6 Hz) and 6.16 (d, j=17.5 Hz, 1H), 4.95-4.81 (m, 2H), 4.62-4.43 (m, 3H), 4.25-4.18 (m, 1H), 3.96-3.83 (m, 1H), 3.38(s) and 3.36 (s, 3H), 2.38-2.21 (m, 4H), 1.75-1.63 (m, 2H), 1.32-1.16 (m, 12H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.04(s),3.97(s).MS(ESI)m/z C ₂₈ H ₄₀ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 636.3; actual measurement value: 636.2.

modification of 2-amino moieties in active Compounds

One of ordinary skill in the art can add substituents to the 2-aminopurine moiety by methods well known to those of ordinary skill in the art. One non-limiting method is provided herein and other methods may be readily employed. Treatment of ((2 r,3r,4r,5 r) -3- (benzoyloxy) -5-bromo-4-fluoro-4-methyltetrahydrofuran-2-yl) benzoate with a mixture of commercially available 2, 6-dichloropurine, base and organic solvent at elevated temperature gives (2 r,3r,4r,5 r) -5- (2, 6-dichloro-9H-purin-9-yl) -2- (benzoyloxymethyl) -4-fluoro-4-methyltetrahydrofuran-3-yl benzoate. In one embodiment, the base is potassium t-butoxide. In one embodiment, the mixture of organic solvents includes t-butanol and acetonitrile. Treatment of the compound (2R, 3R,4R, 5R) -5- (2, 6-dichloro-9H-purin-9-yl) -2- (benzoyloxymethyl) -4-fluoro-4-methyltetrahydrof-N-3-ylbenzoate with an amine, a base and an organic solvent at ambient temperature gives 2-chloro-N ⁶ -substituted purines. In one embodiment, the amine is methylamine. In one embodiment, the base is triethylamine. In one embodimentIn the case, the organic solvent is ethanol. Those skilled in the art will also appreciate that the benzoate groups on the nucleosides should be removed simultaneously when treated with an amine and a base to produce a deprotected furanose moiety. The 2-chloro-N can then be treated with an amine and an organic solvent in a sealed tube at an elevated temperature of about 100deg.C ⁶ Substituted purines giving N according to the invention ² ,N ⁶ -disubstituted purine nucleosides. In one embodiment, the amine is methylamine. In one embodiment, the organic solvent is ethanol. The N-alanine isopropyl ester of the present invention may be treated with a base, ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, and an organic solvent at a reduced temperature ² ,N ⁶ -disubstituted purine nucleosides, yielding compounds of formula I-V. In one embodiment, the base is t-butyl magnesium chloride. In one embodiment, the organic solvent is tetrahydrofuran.

Preparation of stereospecific phosphorus enantiomer

Some of the active compounds described herein have chiral phosphorus moieties. Any of the active compounds described herein can be provided as isolated phosphorus enantiomer forms, e.g., at least 80, 90, 95, or 98% of the R or S enantiomer, using methods known to those skilled in the art. For example, there are numerous publications describing how such compounds can be obtained, including but not limited to column chromatography, such as example 17 below and U.S. Pat. No.8,859,756 to Ross et al; 8,642,756 and 8,333,309.

EXAMPLE 9 separation of stereoisomers of Compound 5

The stereoisomers of Compound 5 were separated on a Phenominex Luna column using the conditions column Phenominex Luna 5 mini-ron C18 (2) 250x 10mm part#OOG-4252-BO

Sample concentration: about 50mg/mL in acetonitrile

Injection volume: 50. Mu.l

Mobile phase A: HPLC grade water

Mobile phase B: HPLC grade acetonitrile.

Flow rate: 5ml/min

UV：283nm

Gradient of

Time	％B
		0	2
40	50
		41	50
41.1	2
		45	2

Run time: 45 minutes

Column temperature: 40 DEG C

A sample chromatogram of a semi-preparative run is shown in fig. 1.

The pooled fractions were analyzed using an analytical column under the following conditions

Column Phenominex Luna 5 mini-ron C18 (2) 250x 2mm part#OOG-4252-BO

Injection volume: 10. Mu.l

Mobile phase A: HPLC grade water

Mobile phase B: HPLC grade acetonitrile.

Flow rate: 0.2ml/min

UV：283nm

Gradient of

Time	％B
		0	2
30	50
		40	50
40.1	2
		45	2

Run time: 45 minutes

Column temperature: 40 DEG C

The combined fractions of each stereoisomer were evaporated to dryness using a rotary evaporator (rotovap) at a bath temperature of 30 ℃. The resulting solid was dissolved in 1ml acetonitrile, transferred to a 1.7ml microcentrifuge tube, and the solvent was evaporated on a vacuum centrifuge at a temperature of 30 ℃. .

The data for the final samples are as follows

1. The first elution peak: compound 5#1 (5-1) (21.7 mgs-97.8% ee).

2. The second elution peak: compound 5#2 (5-2) (13.2 mgs-95.9% ee).

The final weights of the first and second peaks correspond exactly to their percentages in the original mixture (62.2% and 37.8%, respectively).

Stereospecific synthesis of compounds of formulas I-VII

EXAMPLE 10 preparation of (2R, 3R,4R, 5R) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyltetrahydrof-n-3-ol (23)

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyltetrahydrof-n-3-ol (23)

The compound (2 r,3r,4r,5 r) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (benzoyloxymethyl) -4-fluoro-4-methyltetrahydrof-n-3-ylbenzoate 3 (80 g,140 mmol) was added to a solution of trimethylamine in methanol (7 m,800 ml) and stirred overnight at room temperature. The mixture was concentrated and then purified by column chromatography (DCM: meoh=100:1) to give (2 r,3r,4r,5 r) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyl-tetrahydrofuran-3-ol (23) (40 g, 90%).

Preparation of(((S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine ester.

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (4)

To a solution of (2 r,3r,4r,5 r) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyl-tetrahydrofuran-3-ol (2.0 g,1.0 eq) in dioxane (15 mL) was added MeNH ₂ Aqueous solution (5.0 eq). After stirring overnight at room temperature, TLC showed exhaustion of the starting material given. The mixture was concentrated and purified by column chromatography (DCM: meoh=40:1-30:1)The reaction mixture was converted to (2R, 3R,4R, 5R) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (1.6 g, 81.6%) as a white powder.

[M+H] ⁺ ＝313.5

Preparation of((((S) - (2 r,3r,4r,5 r) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine ester.

The compound (2 r,3r,4r,5 r) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (1.47 g,1.0 eq) and PPAL-S (2.35 g,1.1 eq) were dissolved in anhydrous THF (29 mL). After cooling the mixture to-10 ℃, at N ₂ Under an atmosphere (blancet), t-BuMgCl (5.8 mL,1.7M,2.1 eq) was slowly added. After stirring at room temperature for 45 minutes, saturated NH was used ₄ The mixture was quenched with aqueous Cl and extracted with EtOAc (20 mL. Times.3). The combined organic layers were washed with water, brine (30 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography (DCM: meoh=50:1-20:1) to give ((((S) - (2 r,3r,4r,5 r) -5- (2-amino-6- (methylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyl-tetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine ester (1.1 g, 40.3%) as a white powder.

¹ H NMR(400MHz,CD ₃ OD)δ7.81(s,1H),7.33-7.16(m,5H),6.10(d,J＝18.4Hz,1H),4.90-4.84(m,5H),4.55-4.46(m,3H),4.20-4.16(m,1H),3.91-3.87(m,1H),3.30(m,1H),3.03(s,3H),1.30-1.20(m,12H).[M+H] ⁺ ＝582.8.

Preparation of (((S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (25).

Preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol

To a solution of (2 r,3r,4r,5 r) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyl-tetrahydrofuran-3-ol (2.8 g,8 mmol) in dioxane (20 mL) was added aqueous dimethylamine (5 mL). After stirring at room temperature for 3 hours, TLC showed exhaustion of the starting material given. The mixture was concentrated and purified by column chromatography (DCM: meoh=60:1) to give (2 r,3r,4r,5 r) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (2.2 g).

¹ H NMR(400MHz,CD ₃ OD)δ8.08(s,1H),6.13(d,J＝18.0Hz,1H),4.43(dd,J＝9.2,9.2Hz,1H),4.06(d,J＝10.8Hz,2H),3.90(m,1H),3.37(s,3H),3.06(s,3H),1.18(d,J＝22Hz,3H).

Preparation of ((((S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (25)

The compound (2 r,3r,4r,5 r) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (8 g,1.0 eq) and PPAL-S (11.1 g,1 eq) were dissolved in anhydrous THF (100 mL). The mixture was cooled to-5-0deg.C and cooled to N ₂ t-BuMgCl (30.5 mL,1.7M,2.1 eq) was slowly added under the atmosphere. After stirring at room temperature for 2 hours, saturated NH was used ₄ The mixture was quenched with aqueous Cl and extracted with EtOAc (70 ml×3). The combined organic layers were washed with water, brine (30 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating.The crude product was purified by column chromatography (DCM: meoh=50:1) to give ((((S) - (2 r,3r,4r,5 r) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester as a white powder (9.5 g, 65%).

¹ H NMR(400MHz,CD ₃ OD)δ7.81(s,1H),7.35-7.19(m,5H),6.15(d,J＝18.8Hz,1H),4.90(m,1H),4.54-4.49(m,3H),4.22-4.19(m,1H),3.90(m,1H),3.43(s,3H),1.32(d,J＝7.2Hz,3H),1.24-1.17(m,9H). ³¹ P NMR(160MHz,CD ₃ OD)δ3.89。

EXAMPLE 13 preparation of ((((R) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (26)

The compound (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (3 g,1.0 eq) and PPAL-R (4.17 g,1 eq) were dissolved in anhydrous THF (60 mL). The mixture was cooled to-5-0deg.C and cooled to N ₂ t-BuMgCl (11.4 mL,1.7M,2.1 eq) was slowly added under atmosphere. After stirring at room temperature for 16 hours, saturated NH was used ₄ The mixture was quenched with aqueous Cl and extracted with EtOAc (50 ml×3). The combined organic layers were washed with water, brine (30 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography (DCM: meoh=50:1) to give ((((R) - (2R, 3R,4R, 5R) -5- (2-amino-6- (dimethylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester as a white powder (2.2 g, 41%).

¹ H NMR(400MHz,CD ₃ OD)δ7.8(s,1H),7.35-7.29(m,5H),6.18(d,J＝18.8Hz,1H),4.92(m,1H),4.60(m,1H),4.51-4.23(m,3H),3.90(m,1H),3.44(s,6H),1.29(d,J＝6Hz,3H),1.22-1.16(m,10H). ³¹ P NMR(160MHz,CD ₃ OD)δ3.98。

EXAMPLE 14 preparation of ((((S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester

Step 1 preparation of (2R, 3R,4R, 5R) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (8)

Will K ₂ CO ₃ (53 g,500 mmol) was added to an aqueous solution of N-methylcyclopropylamine hydrochloride (100 mL). After stirring at room temperature for 10 minutes, a solution of (2 r,3r,4r,5 r) -5- (2-amino-6-chloro-9H-purin-9-yl) -2- (hydroxymethyl) -4-fluoro-4-methyl-tetrahydrofuran-3-ol (35 g,109 mmol) in dioxane (300 ml) was added. The mixture was stirred at room temperature for 16 hours, HPLC indicated the reaction was complete. The mixture was concentrated and purified by column chromatography (DCM: meoh=60:1) to give (2 r,3r,4r,5 r) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-3-ol (30 g, 82%).

¹ H NMR(400MHz,CD ₃ OD)δ8.16(s,1H),6.17(d,J＝18.0Hz,1H),4.41(dd,J＝9.2,9.2Hz,1H),4.06(m,2H),3.90(m,1H),3.37(s,3H),3.16(m,1H),1.18(d,J＝22.4Hz,3H),0.94(m,2H),0.74(m,2H).[M+H] ⁺ ＝353.2.

Step 2 preparation of (((S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

The compound (2 r,3r,4r,5 r) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (8 g,1.0 eq) and PPAL-S (10.3 g,1 eq) were dissolved in anhydrous THF (100 mL). After cooling the mixture to-5-0deg.C, at N ₂ t-BuMgCl (28 mL,1.7M 2.1 eq) was slowly added under atmosphere. The mixture was stirred at room temperature for 1 hour with saturated NH ₄ The aqueous Cl solution was quenched and extracted with EtOAc (70 mL. Times.3). The combined organic layers were washed with water, brine (30 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography (DCM: meoh=100:1 to 50:1) to give ((((S) - (2 r,3r,4r,5 r) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (9.5 g, 65%) as a white powder.

¹ H NMR(400MHz,CD ₃ OD)δ7.86(s,1H),7.35-7.19(m,5H),6.17(d,J＝19.2Hz,1H),4.91(m,1H),4.52(m,3H),4.21(m,1H),3.93(m,1H),3.35(s,3H),3.16(m,1H),2.0(s,1H),1.26-1.16(m,12H),0.93(m,2H),0.73(m,2H). ³¹ P NMR(160MHz,CD ₃ OD)δ3.90

Preparation of (((R) - (2R, 3R,4R, 5R) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrof-n-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

The compound (2R, 3R,4R, 5R) -5- (2-amino-6- (methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-methyltetrahydrofuran-3-ol (3 g,1.0 eq) and PPAL-R (2.8 g,1 eq) were dissolved in anhydrous THF (60 mL). After cooling the mixture to-5-0deg.C, at N ₂ t-BuMgCl (7.6 mL,1.7M,2.1 eq) was slowly added under atmosphere. The mixture was then stirred at room temperature for 1 hour with saturated NH ₄ The aqueous Cl solution was quenched and extracted with EtOAc (50 mL. Times.3). The combined organic layers were washed with water, brine (30 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography (DCM: meoh=100:1 to 50:1) to give the product as a white powder (3 g, 55%).

¹ H NMR(400MHz,CD ₃ OD)δ7.81(s,1H),7.30-7.25(m,5H),6.16(d,J＝24.8Hz,1H),4.84(m,1H),4.84-4.50(m,3H),4.22-4.19(m,1H),3.88(m,1H),3.33(s,3H),3.14(m,1H),2.0(s,1H),1.28-1.13(m,12H),0.92(m,2H),0.90(m,2H). ³¹ P NMR(160MHz,CD ₃ OD)δ3.99.

EXAMPLE 16 preparation of Compound 32

Step 1. Preparation of compound 29.

To a solution of 6 (3.0 g,1.0 eq) in pyridine (30 mL) was added TIPDSCl at 0deg.C ₂ (4.35 g,1.5 eq). After stirring at room temperature for 4 hours, TLC showed starting material depletion. The mixture was diluted with EtOAc and saturated aqueous NaHCO with 1M HCl ₃ Washing with aqueous solution and brine, passing through anhydrous Na ₂ SO ₄ Drying and concentration gave 29 (6.3 g, 100%) as a yellow oil. .

Step 2. Preparation of compound 30.

Isobutyryl chloride (209 mg,1.5 eq) was added to a mixture of compound 29 (800 mg,1.0 eq), DMAP (16 mg,0.1 eq), pyridine (1.6 mL) and DCM (10 mL) at 0 ℃. After stirring at room temperature for 2 hours, TLC showed starting material depletion. The mixture was quenched with water, 1M aqueous HCl, saturated NaHCO ₃ Washing with aqueous solution and brine, passing through anhydrous Na ₂ SO ₄ Drying and concentrating. Purifying the crude product by column chromatography to obtainTo product 30 (563 mg, 62.3%) as a white oil.

¹ H NMR(400MHz,CDCl ₃ )δ7.98(s,1H),787(s,1H),6.20(d,J＝16.0Hz,1H),4.32-4.07(m,4H),3.50(s,6H),2.3(m,1H),1.29-1.05(m,45H).

Step 3. Preparation of Compound 31.

To a mixture of 30 (560 mg,1.0 eq) in THF (10 mL) at room temperature was added E _t3 N.3HF (706 mg,5 eq) and Et ₃ N (89mg, 10 eq). After stirring at room temperature for 1.5 hours, TLC showed starting material depletion. The mixture was concentrated and purified by column chromatography to give 31 (288 mg, 83%) as a white powder.

¹ H NMR(400MHz,CDCl ₃ )δ7.72(s,1H),5.96(d,J＝44.0Hz,1H),5.22(m,1H),4.13-3.99(m,4H),3.42(s,6H),2.83-2.63(m,2H),1.29-1.17(m,9H).

Step 4. Preparation of compound 32.

Compound 31 (280 mg,1.0 eq) and PPAL-S (320 mg,1 eq) were dissolved in anhydrous THF (10 mL). After cooling the mixture to-5 ℃, at N ₂ t-BuMgCl (0.87 mL,1.7M,2.1 eq) was slowly added under atmosphere. The mixture was stirred at room temperature for 2 hours with saturated NH ₄ The aqueous Cl solution was quenched and extracted with EtOAc (10 mL. Times.3). The combined organic layers were washed with water, brine (20 mL), and dried over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography to give the product as a white powder (260 mg, 50%).

¹ H NMR(400MHz,CD ₃ OD)δ7.98(s,1H),7.25(m,5H),6.23(d,J＝18.8Hz,1H),4.52(m,3H),4.38(m,1H),3.81(m,1H),3.75(m,1H),3.48(s,6H),2.81(m,1H),1.32(m,18H).[M+H] ⁺ ＝666.9.

Example 17 preparation of compound 35.

Step 1. Preparation of compound 33.

AcCl (0.414 g,1.5 eq) was added to a mixture of 29 (2.0 g,1.0 eq), DMAP (0.04 g,0.1 eq), pyridine (4 mL) and DCM (20 mL) at 0deg.C. After stirring at room temperature for 2 hours, TLC showed starting material depletion. The mixture was quenched with water, 1M aqueous HCl, saturated NaHCO ₃ Aqueous solution and then brine, over anhydrous Na ₂ SO ₄ Drying and concentrating. The crude product was purified by column chromatography to give product 33 (1.73 g, 80.8%) as a white oil.

¹ H NMR(400MHz,CDCl ₃ )δ7.99(s,1H),7.74(s,1H),6.20(d,J＝20.0Hz,1H),4.33-4.11(m,4H),3.50(s,6H),2.63(s,3H),2.3(m,1H),1.26-1.05(m,29H).[M+H] ⁺ ＝611.9.

Step 2. Preparation of compound 34.

To a mixture of 33 (1.58 g,1.0 eq) in THF (20 mL) at room temperature was added Et ₃ N.3HF (2.1 g,5 eq) and Et ₃ N (2.6 g,10 eq). After stirring at room temperature for 1.5 hours, TLC showed starting material depletion. The mixture was concentrated and purified by column chromatography to give 34 (782 mg, 82%) as a white powder.

[M+H]+＝369.6.

Step 3. Preparation of compound 35.

Compound 34 (136 mg,1.0 eq) and PPAL-S (184 mg,1.1 eq) were dissolved in anhydrous THF (3 mL). After cooling the mixture to-5 ℃, at N ₂ t-BuMgCl (0.5 mL,1.7M,2.1 eq) was slowly added under the atmosphere. The mixture was stirred at room temperature for 30 min with saturated NH ₄ The aqueous Cl solution was quenched and extracted with EtOAc (10 mL. Times.3). The combined organic layers were washed with water, brine (20 mL), dried over anhydrous and concentrated. Purification by column chromatography (DCM: meoh=50:1-20:1) afforded phosphoramidate 35 (150 mg, 63.8%) as a white powder.

¹ H NMR(400MHz,CD ₃ OD)δ7.81(s,1H),7.35-7.16(m,5H),6.10(d,J＝18.4Hz,1H),4.87(m,1H),4.52-4.46(m,3H),4.21(m,1H),3.91-3.87(m,1H),3.03(s,3H),1.30-1.13(m,12H). ³¹ P NMR(160MHz,CD ₃ OD)δ3.84. ¹⁹ F NMR(376MHz,CD ₃ OD)δ-162.79。

beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-ethynyl-N ⁶ Synthesis of substituted-2, 6-diaminopurine nucleotides

EXAMPLE 18 beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-ethynyl-N ⁶ General synthetic route to substituted 2, 6-diaminopurine nucleotides

Step 1. Preparation of compound 36.

At N ₂ TPDSCl was added dropwise to a solution of 6-chloroguanosine (100 g,332 mmol) in pyridine (400 mL) at-5 to 5℃under an atmosphere ₂ (110 mL,1.05 eq.). After stirring at this temperature for 2 hours, TLC showed starting material depletion. DCM (600 mL) was added, followed by dropwise addition of TMSCl (85 mL,2 eq.) at 0-5 ℃. After stirring at this temperature for 2 hours, TLC showed intermediate depletion.

Isobutyryl chloride is added dropwise at 0-5 ℃. After stirring at this temperature for 2 hours, TLC showed intermediate depletion. Water was added and the contents extracted with DCM. The organic phase was then washed with 0.5N HCl to remove pyridine. .

After washing the pH of the contents to 5 to 6, pTSA.H is added at 0-5 DEG C ₂ O (9.2 g,484.5 mmol). After stirring at this temperature for 1 hour, TLC showed intermediate depletion. Then, add water, and use water, saturated NaHCO ₃ The organic phase was washed with aqueous solution and brine. In the Na way ₂ SO ₄ After drying, the solvent was removed under vacuum. The residue was then purified by column chromatography (PE/ea=100-10/1) to give the product as a pale yellow solid (82 g, 40%).

¹ H NMR(400MHz,DMSO-d ₆ )δ10.88(s,1H),8.55(s,1H),5.91(d,J＝1.6Hz,1H),5.53(d,J＝4.6Hz,1H),4.72–4.58(m,2H),4.16(dd,J＝12.4,4.8Hz,1H),4.00(ddd,J＝7.7,4.8,2.6Hz,1H),3.93(dd,J＝12.4,2.7Hz,1H),2.78(h,J＝6.9Hz,1H),1.26–1.12(m,3H),1.10(d,J＝6.7Hz,6H),1.09–0.88(m,24H)。

Step 2. Preparation of Compound 37.

To a solution of 36 (10.0 g,16.3 mmol) in DCM (100 mL) was added Dess-Martin periodate and the reaction stirred for 12 hours at room temperature. TLC showed starting material depletion. The reaction mixture was then diluted with DCM (200 mL) and saturated Na ₂ S ₂ O ₃ Aqueous solution and brine wash. Then, through Na ₂ SO ₄ The organic phase was dried and concentrated to give crude 37 (12 g) as a pale yellow solid. This crude 53 was used directly in the next step without purification.

Step 3. Preparation of Compound 38.

At N ₂ To a solution of ethynyl trimethylsilane (18.6 mL,142.7 mmol) in THF (240 mL) was added dropwise n-BuLi (46 mL,2.5M,115.0 mmol) at-15-20deg.C under an atmosphere. After stirring for 30 min, the reaction was cooled to-70 ℃ and 37 (crude, 16.3 mmol) in THF (60 mL) was added at that temperature. Then, the content was warmed to 0 ℃. TLC showed starting material depletion. Adding saturated NH ₄ Aqueous Cl and the reaction was extracted three times with EA (100 mL). The organic phases were combined and then washed with brine, followed by further Na ₂ SO ₄ And (5) drying. After concentration in vacuo, the mixture was purified by column chromatography (PE/ea=100->10/1) the residue was purified to give a pale yellow solid (6.0 g, 52%).

Step 4. Preparation of Compound 39.

At N ₂ To a solution of 38 (6.0 g,8.4 mmol) in DCM (240 mL) was added pyridine (4.2 mL,52.9 mmol) under an atmosphere. The reaction was cooled to-70℃and DAST (12 mL,90.4 mmol) was added. Then, the content was warmed to-30 ℃. TLC showed starting material depletion. The reaction was poured onto saturated NaHCO ₃ In aqueous solution, then extracted with DCM (200 mL). The organic phase was washed with brine and dried over Na ₂ SO ₄ And (5) drying. After vacuum drying, column chromatography (PE/ea=100->10/1) the residue was purified to give a pale yellow solid (3.8 g, 63%).

Step 5. Preparation of compound 40.

To a solution of 39 (3.8 g,5.3 mmol) in THF (120 mL) was added AcOH (1.3 g,22 mmol) and TBAF (4.2 g,15.9 mmol) at room temperature. The reaction was stirred at room temperature for 30 minutes. TLC showed starting material depletion. After drying in vacuo, the residue was purified by column chromatography (EA) to give the product as a white solid (2.0 g, 95%).

General procedure for amino substitution and deprotection

To a solution of 40 (350 mg,0.88 mmol) in dioxane (20 mL) was added an aqueous solution of methanol or the corresponding amine (free base or salt such as hydrochloride+diea) at room temperature. The contents were stirred at room temperature for 1-12 hours. TLC showed starting material depletion. After concentration in vacuo, the residue was used directly in the next step without purification. The residue was dissolved in methanol (10 mL). Aqueous NaOH (2.5 n,10 ml) was added. After stirring overnight at room temperature, TLC showed starting material depletion. The pH of the contents was adjusted to 7-8 with 1N HCl. The solution was concentrated and purified by column chromatography (DCM/meoh=100- > 20/1) to give the product as an off-white solid (yield over 2 steps: 40-80%). Table 1 illustrates the structures of compounds 57-63, as well as the corresponding mass spectra and 1H NMR of each compound.

Table 1.

EXAMPLE 19 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6-dimethylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyl-tetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester

i) ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester, tBuMgCl, THF, 0deg.C.

Preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6-dimethylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyl-tetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

To a solution of compound 41 (30 mg,0.09 mmol) in anhydrous THF (2 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 125 μL,0.13 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes and at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (49 mg,0.11 mmol) in anhydrous THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give the product as a white solid (mixture of 2 diastereomers, 12mg,0.02mmol, 24%).

¹ H NMR(300MHz,CD ₃ OD) delta 7.79 (s, 0.45H), 7.77 (s, 0.55H), 7.36-7.14 (m, 5H), 6.28 (d, j=17.4 Hz) and 6.26 (d, j=17.5 Hz, 1H), 5.00-4.44 (m, 5H), 4.23-4.16 (m, 1H), 3.69-3.81 (m, 1H), 3.42 (bs, 3H), 3.40 (bs, 3H), 1.32-1.26 (m, 3H), 1.20-1.15 (m, 6H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.04(s),3.98(s).MS(ESI)m/z C ₂₆ H ₃₄ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 606.2; actual measurement value: 606.2.

preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6-methylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

Preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6-methylamino-9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyl tetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

To a solution of 42 (30 mg,0.09 mmol) in anhydrous THF (2 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1.0M 125 μL in THF, 0.13 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes and then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (49 mg,0.11 mmol) in anhydrous THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give the product as a white solid (mixture of 2 diastereomers, 9mg,0.02mmol, 18%).

¹ H NMR(300MHz,CD ₃ OD) delta 7.81,7.79 (0.9s+0.1s, 1h), 7.36-7.14 (m, 5H), 6.26 (d, j=17.4 hz, 0.1h) and 6.24 (d, j=17.4 hz, 0.9H), 4.93-4.89 (overlapped with H) ₂ O,m,1H),4.80-4.78(m,1H),4.53-4.49(m,2H),4.21-4.18(m,1H),3.95-3.84(m,1H),3.23-3.20(m,1H),3.04(bs,1H),1.31-1.14(m,9H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.06(s),3.97(s).MS(ESI)m/z C ₂₅ H ₃₂ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 592.2; actual measurement value: 592.2.

EXAMPLE 21 preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester

Preparation of ((((R, S) - (2R, 3R,4R, 5R) -5- (2-amino-6- (N-methylcyclopropylamino) -9H-purin-9-yl) -4-fluoro-3-hydroxy-4-ethynyltetrahydrofuran-2-yl) methoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester.

To a solution of compound 43 (40 mg,0.11 mmol) in anhydrous THF (2 mL) at 0deg.C was added dropwise t-butylmagnesium chloride (1.0M in THF, 160 μL,0.16 mmol) over 10 min. The reaction mixture was stirred at 0 ℃ for 15 minutes, then at room temperature for another 15 minutes. The reaction mixture was cooled to 0 ℃ and a solution of ((R, S) - (pentafluorophenoxy) -phenoxy-phosphoryl) -L-alanine isopropyl ester (55 mg,0.12 mmol) in anhydrous THF (2 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 18 hours. With saturated NH ₄ The reaction was quenched with aqueous Cl (4 mL) and extracted with EtOAc (3X 5 mL). The combined organics were purified over Na ₂ SO ₄ Drying and concentrating. The residue was purified by column chromatography (gradient DCM/MeOH 100:0 to 90:10) to give the product as a white solid (mixture of 2 diastereomers, 18mg,0.03mmol, 26%).

¹ H NMR(300MHz,CD ₃ OD) delta 7.84,7.82 (s+s, 1H), 7.35-7.14 (m, 5H), 6.30 (d, j=17.4 Hz) and 6.26 (d, j=17.6 Hz, 1H), 4.99-4.89 (with H) ₂ O overlap, m, 1H), 4.82-4.69 (m, 1H), 4.59-4.46 (m, 2H), 4.21 (m, 1H), 3.96-3.82 (m, 1H), 3.24-3.22 (m, 1H), 3.17-3.11 (m, 1H) 1.31-1.26 (m, 3H), 1.20-1.15 (m, 6H), 0.93-0.89 (m, 2H), 0.75-0.68 (m, 2H). ³¹ P NMR(121MHz,CD ₃ OD)δ4.06(s),3.98(s).MS(ESI)m/z C ₂₈ H ₃₆ FN ₇ O ₇ P[M+H] ⁺ Is the theoretical value of (3): 632.2; actual measurement value: 632.2.

EXAMPLE 22 preparation of PPAL-S

Step 1 preparation of racemic PPAL

To a stirred solution of phenyl dichlorophosphate (250 g) in EtOAc (800 mL) at-10deg.C was added isopropyl L-alaninate (200 g) in triethylamine (120 g). The reaction was stirred at-10℃for 1 hour. 2,3,4,5, 6-pentafluorophenol (220 g) as a compound in triethylamine (120 g) and EtOAc (400 mL) was added at-5℃and stirred at that temperature for 0.5 hours. The reaction mixture was warmed to 25 ℃ and stirred at that temperature for 2 hours. The solution was filtered and washed with EtOAc (2×200 mL) and the combined organic phases were evaporated in vacuo to give PPAL-RS (racemate) as a solid.

Preparation of PPAL-RS

To a stirred solution of PPAL-RS in EtOAc (200 mL) and n-heptane (1.4L) was added 2,3,4,5, 6-pentafluorophenol (10.1 g) in triethylamine and stirring continued for about 4-8 hours. After less than 0.5% of the R-isomer of the solid, the solid was filtered. The solid was dissolved in EtOAc (4L), washed with water (2X 100 mL), brine (1L) and dried over anhydrous Na ₂ SO ₄ Drying and filtering. The solvent was removed under vacuum to give PPAL-S (350 g).

¹ H NMR(400MHz,DMSO-d6)δ＝7.42–7.40(m,2H),7.24–7.22(m,3H),6.87(dd,J＝14.1,9.9Hz,1H),4.90–4.84(m,1H),3.94–3.88(m,1H),1.27(dd,J＝7.1,1.1Hz,3H),1.15(dd,J＝6.2,1.2Hz,6H)ppm.. ¹³ P NMR(160MHz,DMSO-d6)δ＝0.37ppm.

EXAMPLE 23 preparation of PPAL-R

To a three-necked round bottom flask equipped with a mechanical stirrer were added phenyl dichlorophosphate (189.6 g,0.90 mol) and anhydrous EtOAc (750 mL). The solution was cooled to-10 ℃ under nitrogen. L-alanine isopropyl ester (118 g,0.90 mmol) and triethylamine (100 g,1.1 eq) were added to the above solution. A pre-cooled (below 10 ℃) mixture of 2,3,4,5, 6-pentafluorophenol (165 g,1 eq) and triethylamine (90.5 g,1 eq) in EtOAc (300 mL) was added to the mixture via an addition funnel at-5 ℃ and the resulting mixture was stirred at 20-25 ℃ for 1 hour. The white precipitate (tea.hcl) was filtered off and washed with EtOAc. The filtrate was concentrated under reduced pressure to give about 280g of PPAL-RS as a white solid (S/R=1/1). PPAL-RS (280 g) was triturated in 300mL of heptane/EtOAc (20:1) for 5 minutes at room temperature. The white suspension was filtered and the solid was washed with a heptane/EtOAc (20:1) mixture. The filtrate was cooled to 8 ℃ and the solid was collected by filtration. Crude PPAL-R (10 g) was obtained, 95% chiral purity. The crude product was purified according to the procedure described above. PPAL-R (5 g), NLT 98% chiral purity was obtained.

¹ H NMR(400MHz,DMSO-d ₆ )δ＝7.43–7.39(m,2H),7.27–7.22(m,3H),6.87(dd,J＝14.1,9.9Hz,1H),4.89–4.85(m,1H),3.95–3.90(m,1H),1.27(dd,J＝7.1,1.1Hz,3H),1.14(dd,J＝6.2,1.2Hz,6H). ¹³ P NMR(160MHz,DMSO-d ₆ )δ＝0.35.

Example 24: preparation of compound 52: .

Step 1. Preparation of Compound 49.

To a solution of 48 (1.81 g,3.23 mmol) in dioxane (18 mL) was added 40% CH ₃ NH ₂ Aqueous solution (16.2 mmol). The reaction was stirred at 40℃for 2 hours. The mixture was concentrated, diluted with EtOAc (50 mL), washed with water and brine. The organic layer was treated with anhydrous Na ₂ SO ₄ Dried, filtered and concentrated to give 49 (1.66 g, 92%) as a white solid. ).

Step 2. Preparation of compound 50.

To a solution of 49 (1.34 g,2.42 mmol) and 1-methylimidazole (794 mg,9.68 mmol) in DCM (14 mL) was slowly added amyl chloroformate (547 mg,3.63 mmol) at 0deg.C. The reaction was stirred at room temperature overnight. The mixture was concentrated and purified by column chromatography (PE: etoac=5:1-2:1) to give 50 (1.01 g, 62%) as a white solid. ¹ HNMR(400MHz,DMSO)δ7.96(s,1H),6.73(s,1H),6.06-6.10(d,J＝16.0Hz,1H),4.09-4.30(m,2H),3.97-4.09(m,4H),3.28(s,3H),1.39-1.46(m,2H),1.0-1.2(m,35H),0.73-0.76(t,J＝8.0Hz,3H)。

Step 3 preparation of Compound 51

Et is added to a solution of 50 (1.00 g,1.5 mmol) in THF (11 mL) at 0deg.C ₃ N (2.0 mL,15 mmol) and Et ₃ N.3HF (1.21 g,7.5 mmol). The reaction was stirred at room temperature for 1.5 hours. The mixture was concentrated and purified by column chromatography (MeOH: ch2cl2=50:1) to give 75 (460 mg, 72.2%) as a white powder. .

Step 4 preparation of Compound 52

At N ₂ To a solution of 51 (460 mg,1.08 mmol) and PPAL-S (538 mg,1.19 mmol) in anhydrous THF (9 mL) was slowly added t-BuMgCl (2.27 mmol) at 5-10deg.C. The reaction was stirred at room temperature for 40 minutes. With saturated NH ₄ The mixture was quenched with aqueous Cl, extracted with EtOAc and concentrated with 5% K ₂ CO ₃ Washing with aqueous solution and brine, washing with anhydrous Na ₂ SO ₄ Dried, filtered and concentrated. By column chromatography (CH ₂ Cl ₂ Meoh=15:1) to give 52 (280 mg, 37.3%) as a white solid. ¹ H NMR(400MHz,DMSO)δ8.12(s,1H),7.34-7.38(m,2H),7.18-7.23(m,3H),6.74(s,2H),6.11-6.16(d,J＝16.0Hz,1H),5.99-6.05(m,1H),5.84(m,1H),4.77-4.81(m,1H),4.30-4.41(m,3H),4.03-4.11(m,3H),3.78-3.80(m,1H),3.3(s,3H),1.44-1.51(m,2H),1.00-1.21(m,16H),0.76-0.80(t,J＝8.0Hz,3H).[M+H] ⁺ ＝696.6.

Example 25 preparation of Compound 56.

Step 1 preparation of Compound 48

To a solution of 23 (600 mg,1 eq) in pyridine (30 mL) was added TIPDSCl at 0deg.C ₂ (1.5 eq). The resulting solution was allowed to stand at room temperature for 2 hours. The mixture was quenched with ice water and extracted with EtOAc. The organic layer was washed with 1M aqueous HCl, saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and concentrated to give a crude residue. The residue was purified by chromatography (MeOH: ch2cl2=1:50) to give 48 (998 mg, 94.4%) as a white solid foam.

Step 2. Preparation of Compound 53.

A mixture of 48 (800 mg,1 eq), pyridine (3.2 mL), DMAP (34.9 mg,0.2 eq) in DCM (20 mL) was stirred at room temperature. N-amyl chloroformate (3.2 mL) was added dropwise at 0 ℃ and the mixture was stirred at room temperature for 1 day. The organic layer was washed with 1M aqueous HCl, saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, and evaporated in vacuo. The residue was purified by silica gel chromatography (MeOH: ch2cl2=1:50) to give 53 as a white solid foam (255 mg, 26%). .

Step 3. Preparation of compound 54.

To a solution of 53 (270 mg,1 eq) in 1, 4-dioxane (10 mL) was added dropwise 40% CH ₃ NH ₂ Aqueous solution (225.7 mg5 eq). The mixture was stirred at room temperature for 2 hours and then concentrated in vacuo. The residue was chromatographed on silica gel (methanol: dichloromethane=1:40) to give 54 (220 mg, 81.7%) as a white solid foam. .

Step 4. Preparation of compound 55.

Triethylamine (1011.9 mg,10 eq) and Et ₃ N.3HF (806.05 mg,5 eq) was added to an ice-cooled solution of 54 (668 mg,1 eq) in THF (10 mL) and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated and purified on silica gel (MeOH: CH ₂ Cl ₂ =1:30) to give 55 (492 mg, 84%) as a white solid foam. .

Step 5 preparation of Compound 56

To a mixture of 55 (113 mg,1 eq) and PPAL-S (120 mg,1 eq) in THF (4 mL) was added dropwise 1.7. 1.7M t-BuMgCl in THF (0.327 mL,2.1 eq) at-10 ℃. The mixture was stirred at room temperature for 1 hour, then saturated NH ₄ The aqueous Cl solution was quenched. The aqueous phase was extracted with EtOAc and the organic phase was washed with brine, dried and concentrated to give a crude residue. The residue was flash chromatographed to give 56 (126 mg, 68.5%) as a white solid. .

¹ H NMR(400MHz,DMSO)δ8.00(s,1H),7.10-7.45(m,5H),6.15-6.20(d,J＝20.0Hz,1H),5.00-5.25(s,1H),4.80-4.86(m,1H),4.45-4.70(m,2H),4.12-4.19(m,3H),3.80-3.85(m,1H),3.04(s,3H),1.60-1.75(m,2H),1.10-1.40(m,16H),0.76-0.80(t,J＝8.0Hz,3H).

³¹ P NMR(160MHz,DMSO)δ3.57.[M+H] ⁺ ＝696.5.

Example 26 preparation of Compound 60.

Step 1. Preparation of compound 57.

At 5.+ -. 5 ℃ to 6 (20 g,1 eq) in CH ₃ Imidazole (16.6 g), TIPDSCl were added sequentially to a solution in CN (100 mL) ₂ (28.9 g,1.5 eq). Will beThe resulting solution was left to stand at room temperature for 4 hours. The mixture was quenched with ice water and extracted with EtOAc. The organic layer was washed with water, a saturated aqueous sodium hydrogencarbonate solution and a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to give a crude residue (32 g). .

Step 2 preparation of Compound 58

To a solution of 57 (9.8 g,1 eq) in THF (4 mL) was added dropwise 1.7-M t-BuMgCl (50 mL,4.8 eq) in THF at 0-5 ℃. The mixture was stirred at room temperature for 0.5 hours, and n-amyl chloroformate (2.7 g,1.05 eq) was slowly added. The mixture was stirred at 0-5℃for 3-4 hours. With saturated NH ₄ The mixture was quenched with aqueous Cl. The aqueous phase was extracted with EtOAc (200 mL) and the organic phase was washed with brine, dried and concentrated to give 58 (10.7 g) as an oil.

Step 3 preparation of Compound 59

Triethylamine (10.119 g) and Et ₃ N.3HF (8.6 g,5 eq) was added to an ice-cooled solution of 58 (7.3 g,1 eq) in THF (100 mL) and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated and chromatographed on silica gel (MeOH: ch2cl2=1:30) to give 59 (4.3 g, 91%) as a white solid.

Step 4. Preparation of compound 60.

To a mixture of 59 (2 g,1 eq) and PPAL-S (2.3 g,1.1 eq) in THF (40 mL) was added dropwise 1.7-M t-BuMgCl in THF (5.6 mL,2.1 eq) at-5 ℃. The mixture was stirred at-20.+ -. 5 ℃ for 1 hour, then saturated with NH ₄ The aqueous Cl solution was quenched. Extraction of aqueous phase with EtOAcThe organic phase was washed with brine, dried and concentrated to give a crude residue. The residue was flash chromatographed to give 60 (1.5 g, 47%) as a white solid.

¹ H NMR(400MHz,CD ₃ OD)δ7.9(s,1H),7.1～7.2(m,5H),6.2(d,J＝20Hz,1H),5.1(br,1H),4.84(m,1H),4.49(m,,2H),4.16(m,1H),4.13(m,2H),3.86(m,1H),3.45(br,6H),1.70(m,2H),1.26(m,4H),1.20(m,6H),1.14(m,6H),0.93(m,3H).[M+H] ⁺ ＝710.5.

Biological data

Example 27 test methods and additional biological data

Huh-7luc/neo ET cells harboring a bicistronic (distructive) HCV genotype 1b luciferase receptor replicon were expressed at 7.5X10 ³ Cells/ml were plated in 96-well plates in duplicate for parallel determination of antiviral efficacy (EC ₅₀ ) And Cytotoxicity (TC) ₅₀ ). The plates were incubated for 24 hours before the compounds were added. Six series of half-log dilutions of the test preparation (high test concentration of 100.0. Mu.M or high test concentration of 1.0. Mu.M) and human interferon-. Alpha.2b (high test concentration of 10.0U/ml) were prepared in the medium of the cell culture and added to the cultured cell wells, each dilution in triplicate. Six wells in the test plate received medium alone as untreated controls. After 72 hours of incubation in the presence of the compound, one of the plates was used to determine cytotoxicity by staining with XTT and the antiviral efficacy of the other plate by determining luciferase receptor. Cytotoxicity and efficacy data were collected and introduced into a set Excel workbook for determination of TC ₅₀ And EC (EC) ₅₀ Values. The data for the compounds of formulas I-VII are set forth in Table 7 below. In addition, FIG. 2 shows HCV replication inhibition curves for compound 5-2 and Sofosbuvir. As can be seen in FIG. 2, compound 5-2 has EC ₅₀ =4 nM, whereas sofosbuvir has EC ₅₀ =53 nM. The y-axis is the percent of viral control and the x-axis is drug concentration (μm). Figure 3 shows HCV replication inhibition curves for compound 25 and sofosbuvir. Compound 25 has EC ₅₀ Sofosbuvir with EC =4 nM ₅₀ =53 nM. The y-axis is the percent of viral control and the x-axis is drug concentration (μm). Drawing of the figure4 shows a comparison of the anti-HCV activity of compounds 5-2, 25, 27 and sofosbuvir in an in-batch test. The y-axis is the percent of viral control and the x-axis is drug concentration (μm).

The relative replication sensitivity of various patient-derived HCV genotypes including wild-type and resistance-related mutants to the test compounds was determined using them. Replicon Resistance Test Vectors (RTV) comprising NS5B genomic regions were prepared using viral RNAs isolated from plasma of HCV patients. Each NS5B region was amplified by reverse transcription polymerase chain reaction and cloned into HCV replicon RTV and then transferred into Huh-7 cells by electroporation. After incubation for 72-96 hours in the absence and presence of serial dilutions of test compound, viral replication was measured by luciferase activity and 50% inhibition concentration (IC ₅₀ Values).

Table 2 shows the IC of compounds 25, 27, 5-2 and Sofosbuvir against various clinical isolates including wild-type and resistance-associated mutants ₅₀ And IC ₉₅ Values. .

For HCV replication, all compounds were more potent than solfebuibd Wei Xianzhu, neither 25, 27 nor 5-2 compounds showed any evidence of cross resistance to the L159F, L F and S282T and C316N mutants. .

TABLE 2 antiviral Activity of test compounds in patient-derived HCV genotypes

Transient transfection assays were performed to determine the sensitivity of the HCV wild-type S282T mutant to the compounds tested. Huh-7 cells were electroporated in the presence of RNA transcribed from the wild type or an S282T HCV replicon plasmid from the T7 promoter. Transfected cells were grown at 7.5X10 ³ Cells/wells were inoculated into Dulbecco's modified Eagle's medium in 96-well plates. At 24 hours of cultureThereafter, the medium is removed and replaced with fresh medium containing no or varying concentrations of the test compound. After an additional 96 hours incubation, anti-HCV activity was measured by luciferase endpoint using the briitelite Plus fluorescent reporter kit (Perkin Elmer, shelton, CT). Two plates were treated in parallel and incubated in parallel and cytotoxicity was assessed by XTT staining with tetrazolium dye.

Table 3 reports the ICs of compounds 25, 27, 5-3 and Sofosbuvir against the HCV wild type and S282T replicon ₅₀ And IC ₉₅ Values.

For HCV replication, all compounds were more potent than solfebuibd Wei Xianzhu, neither the 25, 27 nor 5-2 compounds showed any evidence of cross resistance to the S282T mutant.

TABLE 3 antiviral Activity of test compounds in a transient infection assay for HCV

The stability of the selected compounds in fresh human whole blood and in human liver S9 fractions was determined in cultures containing 10 μm of the test compound. After incubation for 0, 30, 60 minutes and up to 120 minutes, aliquots were removed and immediately extracted with 3 volumes of ice-cold methanol/acetonitrile (1:1, v/v). The extracts were centrifuged and the supernatant analyzed by LC-MS/MS for unchanged concentrations of test compound and possible metabolites.

Figure 5 shows the excellent stability of compound 5-2 and all 2-amino derivatives in human blood.

Interestingly, FIG. 6 shows 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N using the human liver S9 fraction ² -methyl-N ⁶ -methyl-2, 6-diaminopurine nucleoside phosphoramidate to 2' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-N ⁶ In vitro time course dealkylation of methyl-2, 6-diaminopurine nucleoside phosphoramidates And (5) carrying out glycosylation. Moreover, an unexpected, faster and more extensive (extension) cleavage rate of the carbamate moiety by the human liver S9 fraction was observed compared to compound 5-2 and other 2-amino derivatives (fig. 7).

EXAMPLE 28 HCV (gt 1B) NS5B polymerase assay

Inhibition of HCV (gt 1B) NS5B polymerase was determined in triplicate by measuring the repolymerization in a reaction mixture comprising TA in serial dilutions, in vitro transcriptome RNA complementary to the 3' utr region of the HCV (-) strand, a polymerase, radiolabeled ribonucleotides, 250 μm non-competitive rtp and 1 μm competitive rtp. From the resulting inhibition curve, the concentration of TA (IC) yielding 50% inhibition was determined ₅₀ )。.

EXAMPLE 29 human myeloid progenitor cell assay

Fresh human bone marrow progenitor cells (Invitrogen) suspended in BFU-E or GM-CSF specific medium were treated at 10 ⁵ Individual cells/well were added to serial dilutions of TA in 6-well plates in triplicate. After 14 days of incubation, CC was determined using colony count ₅₀ Values. BFU-E colonies were confirmed using the benzidine (benzodene) technique.

Compounds 25, 27 and 5-2 showed no cytotoxicity to bone marrow stem cells in vitro.

Example 30 iPS cardiomyocyte assay

iPS cardiomyocytes (Cellular Dynamics) were cultured at 1.5X10 ⁴ Individual cells/wells were seeded in microliter plates. After 48 hours of culture, the cells were washed and maintenance medium containing serial dilutions of TA was added in triplicate. After 3 days of further culture, cell viability was measured by staining with XTT and CC was calculated ₅₀ Values. .

Compounds 25, 27 and 5-2 showed no cytotoxicity to iPS cardiomyocytes in vitro.

EXAMPLE 31 human DNA polymerase assay

In serial dilutions of TA, 0.05mM dCTP, dTTP and dATP, 10. Mu. Ci [ ³² P]Determination of human DNA polymerases α, β and γ in a reaction mixture of- α -dGTP (800 Ci/mmol), 20 μg of activated calf thymus DNA and another reagent specific for each PolymerInhibition of (CHIMERx) was performed in triplicate. After 30 minutes of incubation, [ alpha ] ³² P]GTP binding, calculation of IC using the resulting culture curve ₅₀ Values.

None of the triphosphates, β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-guanine triphosphates, nor the triphosphates analogues of compounds 25, 27 and 5-2 inhibit human DNA polymerase α, β or γ. .

EXAMPLE 32 human hepatocyte Co-culture

Measurement of micro-patterned human hepatocyte co-culture by micro-templateHepregen Corporation) to assess cytotoxicity and hepatocyte health status in triplicate, human hepatocyte co-cultures of the micro-templates were prepared by seeding cryopreserved female human hepatocytes (single donor) and 3t3 J2 mouse fibroblasts in microtiter plates according to the method established by Hepregen. Every 2 or 3 days, the medium of the culture was replaced with fresh medium containing TA, test preparation (0, 1, 10 or 30 μm) until day 16. ALT and urea content in spent medium was determined on days 2, 5, 7, 9, 12, 16 and 21, and albumin content was determined on days 2, 5, 7 and 9. Cellular ATP levels were measured on days 9 and 21. The ATP signal of the matrix-only control culture (murine 3T3 fibroblasts) was subtracted from the ATP signal of the human HepatoPac co-culture to give a hepatocyte-specific effect. See tables 4, 5 and 6 below. .

When human hepatocytes co-cultured with the micro-templates were cultured for up to 12 days, compound 5-2 at a concentration of up to 30 μm showed no signs of cytotoxicity as measured by ALT leakage, albumin secretion, urea production and cellular ATP content. The smaller cytotoxicity indications detected with prolonged exposure (up to 21 days of incubation) were significantly smaller than those observed with sofosbuvir. See tables 4, 5 and 6 below.

INX-189 was highly cytotoxic to human co-cultured hepatocytes, and all measurements showed reduced albumin secretion and cytotoxicity as early as day 2. Under the same conditions, sofosbub Wei Xianshi is more cytotoxic than AT-511.

TABLE 4 influence of test preparations on cellular ATP concentration

TABLE 5 influence of test preparations on albumin secretion

TABLE 6 influence of test preparations on albumin secretion

EXAMPLE 33 metabolism Studies

Metabolism of compounds 25, 27 and 5-2 was studied at 10 μm concentrations in fresh primary cultures of human, dog and mouse hepatocytes. Hepatocytes from humans (XenoTech, mixed sex, pooled from 10 donors), male beagle dogs (BioreclamationIVT) and male ICR/CD-1 mice (BioreclamationIVT, 8 donors) plated in matrigel-coated 6-well plates were cultured with 10. Mu.M TA single (single). After 2, 4, 6, 8 or 24 hours, intracellular levels of the nucleotide prodrugs and their possible metabolites (prodrugs, monophosphates, triphosphates and nucleosides) were quantified by LC-MS/MS. Concentrations below the lower limit of quantitation (prodrug, monophosphate and nucleoside 1.5 pmol/10) were extrapolated from the standard curve ⁶ Cells, and the triphosphate is 12pmol/10 ⁶ Cells).

The compound β -D-2' -deoxy-2 ' - α -fluoro-2 ' - β -methyl-guanine triphosphate is the main metabolite of compounds 25, 27 and 5-2 observed in cultured human hepatocytes and is a potent inhibitor of HCV (gt 1B) NS5B polymerase, IC ₅₀ 0.15. Mu.M.

Figure 8 shows the major compound 25 metabolites in human hepatocytes.

Figure 9 shows the major compound 27 metabolites in human hepatocytes.

FIG. 10 shows the major compound 5-2 metabolite in human hepatocytes.

FIG. 11 shows the activation pathways for compounds 25, 27 and 5-2. As can be seen, compounds 25, 27 and 5-2 are converted to their corresponding monophosphate analogs, which are then metabolized to the common MP analogs; beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-guanine monophosphate (compound 61). The monophosphate is then stepwise phosphorylated to the active triphosphate: beta-D-2 ' -deoxy-2 ' -alpha-fluoro-2 ' -beta-methyl-guanine triphosphate (compound 62). .

EXAMPLE 34 control

In the above examples, INX-189 (INX-08189/BMS-986094) and Sofosbuvir were used as controls.

Both of the most potent nucleotide prodrug compounds 25 and 27 showed excellent selectivity for CC in Huh-7 cells, human bone marrow stem cells and human cardiac muscle cells ₅₀ The values are all greater than 100. Mu.M. At concentrations of up to 100 μm, no inhibition of human DNA polymerase α, β or γ was observed in all host cell lines, no activity and no toxicity to other RNA or DNA viruses.

Table 7 shows compounds and ECs tested in HCV replicon assay ₅₀ /EC ₉₅ (mu M) and CC ₅₀ Table of (μm) results.

Table 7. Replicon assay results for test compounds.

beta-D-2 ' -D-2' -alpha-fluoro-2 ' -beta-C-substituted-2-modified-N as described herein ⁶ The substituted purine nucleotides show significant activity against HCV virus. The desired relative activity of the compounds according to the invention is determined using well known and conventional assays described in the literature.

For example, anti-HCV activity and cytotoxicity of compounds can be measured in Huh7 ET cells with an HCV subgenomic RNA replicon assay system. (see Korba, et al, antiviral Research 2008,77,56). The results can be summarized by comparison with the positive control 2 '-C-Me-cytosine {2' -C-Me-C } (Pierra, et al, journal of Medicinal Chemistry 2006,49,6614).

Another in vitro assay for anti-hepatitis C virus activity is described in Stuyver et al, U.S. Pat. No.7,718,790 to Pharmasset, inc.

The present specification has been described with reference to embodiments of the application. Those of ordinary skill in the art, in view of the teachings herein, will be able to modify the application according to the intended purpose and such modifications are considered to be within the scope of the application.

The application also relates to the following items.

1. A compound of formula I:

wherein:

y is NR ¹ R ² ；

R ¹ Is C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CH ₂ F、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ 、CF ₂ CH ₃ And CF (compact F) ₂ CF ₃ )、C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl), -OR ²⁵ 、-C(O)R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ 、-C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ Each of which may be optionally substituted;

R ³ And R is ⁴ Together with the oxygen to which they are bound, may form a 3',5' -cyclic prodrug;

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl;

R ²⁸ is C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted.

2. The compound of item 1, wherein

R ⁴ Is a stable phosphate prodrug, and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

3. The compound of item 2, wherein

R ⁴ Is phosphoramidate, and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

4. The compound of item 2, wherein

Y is NR ¹ R ² The method comprises the steps of carrying out a first treatment on the surface of the And

R ¹ is methyl, R ² Is hydrogen and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

5. The compound of item 2, wherein

R ¹ is methyl, R ² Is methyl, and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

6. The compound of item 2, wherein

R ¹ is methyl, R ² Is cyclopropyl and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

7. The compound of item 1, wherein R ¹² Is CH ₃ 。

8. The compound of item 1, wherein R ¹² Is ethynyl.

9. The compound of the formula or the pharmaceutically acceptable salt thereof

Wherein:

R ⁷ is hydrogen, C _1-6 An alkyl group; c (C) _3-7 Cycloalkyl; heteroaryl, heterocycle, or aryl, wherein phenyl or naphthyl is optionally substituted with: c (C) _1-6 Alkyl, C _2-6 Alkenyl, C _2-6 Alkynyl, C _1-6 Alkoxy, F, cl, br, I, nitro, cyano, C _1-6 Haloalkyl, - (CH) ₂ ) _1-6 COOH、-(CH ₂ ) _1-6 COOC _1-6 Alkyl, -N (R) ⁷ ') ₂ 、C _1-6 Amido, NHSO ₂ C _1-6 Alkyl, -SO ₂ N(R ⁷ ') ₂ 、COR ⁷ "sum-SO ₂ C _1-6 An alkyl group; (R) ⁷ ' independently hydrogen or C _1-6 An alkyl group; r is R ⁷ "is-OR ¹¹ or-N (R) ⁷ ) ₂ )；

R ⁸ Is hydrogen, C _1-6 Alkyl, or R ^9a Or R is ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms; wherein n is 2 to4；

R ^9a And R is ^9b The method comprises the following steps: (i) Independently selected from hydrogen, C _1-6 Alkyl, cycloalkyl, - (CH) ₂ ) _c (NR ⁹ ') ₂ 、C _1-6 Hydroxyalkyl, -CH ₂ SH、-(CH ₂ ) ₂ S(O)(Me、-(CH ₂ ) ₃ NHC(＝NH)NH ₂ (lH-indol-3-yl) methyl, (1H-imidazol-4-yl) methyl, - (CH) ₂ ) _c COR ⁹ ", aryl and aryl (C) _1-3 Alkyl) -, which may optionally be selected from hydroxy, C _1-6 Alkyl, C _1-6 Groups substituted with alkoxy, halogen, nitro and cyano; (ii) R is R ^9a And R is ^9b Are all C _1-6 An alkyl group; (iii) R is R ^9a And R is ^9b Together are (CH) ₂ ) _r Thereby forming a spiro ring; (iv) R is R ^9a Is hydrogen and R ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms, (v) R ^9b Is hydrogen and R ^9a And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising contiguous N and C atoms, wherein C is 1 to 6, N is 2 to 4, R is 2 to 5, and wherein R ⁹ ' independently hydrogen or C _1-6 Alkyl, and R ⁹ "is-OR ¹¹ or-N (R) ¹¹ ') ₂ )；(vi)R ^9a Is hydrogen and R ^9b Is hydrogen, CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower cycloalkyl; or (vii) R ^9a Is CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower cycloalkyl, and R ^9b Is hydrogen;

R ¹⁰ is hydrogen, C optionally substituted by alkoxy, di (lower alkyl) -amino or halogen _1-6 Alkyl, C _1-6 Haloalkyl, C _3-7 Cycloalkyl, heterocycloalkyl, aminoacyl, aryl such as phenyl, heteroaryl such as pyridyl, substituted aryl or substituted heteroaryl;

R ¹¹ is optionally substituted C _1-6 Alkyl, optionally substituted cycloalkyl; optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl or optionally substituted acyl groups including, but not limited to, C (O) (C _1-6 An alkyl group); and Y, R ³ And R is ¹² As defined in item 1.

10. The compound of item 9, wherein

Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen or methyl, and R ¹² Is CH ₃ 。

11. The compound of item 9, wherein:

y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl or hydrogen, R ³ Is hydrogen, R ⁷ Is phenyl, R ⁸ Is hydrogen, R ^9a Is hydrogen, R ^9b Is methyl, R ¹⁰ Is isopropyl and R ¹² Is CH ₃ 。

12. The compound of item 9, wherein

Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl and R ¹² Is CH ₃ 。

13. The compound of item 9, wherein

Y is NR ¹ R ² ，R ¹ Is CH ₃ ，R ² Is C (O) OR ³ ，R ³ Is hydrogen and R ¹² Is CH ₃ 。

14. A compound of formula II:

wherein:

y is NR ¹ R ² ；

R ² is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propylIsopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the And wherein R is ¹ And R is ² At least one of them is methyl, CH ₂ F、CHF ₂ Or CF (CF) ₃ ；

R ^3A May be selected from O-, OH, -O-optionally substituted aryl, -O-optionally substituted heteroaryl or optionally substituted heterocyclyl;

R ^3C is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, -O-alkenyl, -O-alkynyl, -O- (C) ₀ -C ₂ ) (cycloalkyl), -O- (C) ₀ -C ₂ ) (heterocyclyl), -O- (C) ₀ -C ₂ ) (aryl) -O- (C) ₀ -C ₂ ) (heteroaryl), -S-alkyl,-S-alkenyl, -S-alkynyl, -S- (C) ₀ -C ₂ ) (cycloalkyl), -S- (C) ₀ -C ₂ ) (heterocyclyl), -S- (C) ₀ -C ₂ ) (aryl) or-S- (C) ₀ -C ₂ ) (heteroaryl), each of which may be optionally substituted;

R ⁵ is C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ),C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle), - (C ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl), -OR ²⁵ 、-C(O)R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ Each of which may be optionally substituted;

R ⁶ is hydrogen, optionally substituted C ₁ -C ₅ Alkyl (including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl), C ₁ -C ₅ Haloalkyl (including CHF) ₂ 、CHF ₂ 、CF ₃ 、CH ₂ CF ₃ And CF (compact F) ₂ CF ₃ ) Optionally substituted- (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heterocycle), optionally substituted- (C) ₀ -C ₂ Alkyl) (aryl), optionally substituted- (C) ₀ -C ₂ Alkyl) (heteroaryl), -C (O) R ^3C (comprising-C (O) CH ₃ 、–C(O)CH ₂ CH ₃ -C(O)CH(CH ₃ ) ₂ 、-C(O)OCH ₃ 、-C(O)OC ₂ H ₅ 、-C(O)OC ₃ H ₇ 、-C(O)OC ₄ H ₉ and-C (O) OC ₅ H ₁₁ )、-C(S)R ^3D or-SO ₂ R ²⁸ The method comprises the steps of carrying out a first treatment on the surface of the Or R is ⁵ And R is ⁶ Together with the nitrogen to which they are bound may form a heterocyclic ring;

R ¹² is CH ₃ 、CH ₂ F、CHF ₂ 、CF ₃ Or ethynyl;

R ²² f, cl, br, CN, N of a shape of F, cl, br, CN, N ₃ 、C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl); ONHC (=o) OR ²³ 、-NHOR ²⁴ 、-OR ²⁵ 、-SR ²⁵ 、-NH(CH ₂ ) _1-4 N(R ²⁶ ) ₂ 、-NHNHR ²⁶ 、-N＝NR ²⁷ 、-NHC(O)NHNHR ²⁷ 、-NHC(S)NHNHR ²⁷ 、-C(O)NHNHR ²⁷ 、-NR ²⁷ SO ₂ R ²⁸ 、-SO ₂ NR ²⁷ R ²⁹ 、-C(O)NR ²⁷ R ²⁹ 、-CO ₂ R ²⁹ 、-SO ₂ R ²⁹ ,-P(O)H(OR ²⁹ )、-P(O)(OR ²⁹ )(OR ³⁰ )、-P(O)(OR ²⁹ )(NR ²⁹ R ³⁰ ) or-NR ⁵ R ⁶ ；

R ²⁶ independently selected from hydrogen, C ₁ -C ₆ Alkyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (heterocycle)、-(C ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen;

R ²⁷ hydrogen or optionally substituted C ₁ -C ₆ An alkyl group;

R ²⁸ is C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted;

R ²⁹ is hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) or- (C ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen; or (b)

R ³⁰ Is hydrogen, C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl), or- (C) ₀ -C ₂ Alkyl) (heteroaryl), each of which may be optionally substituted with the exception of hydrogen; or (b)

R ²⁹ And R is ³⁰ May be taken together to form a heterocyclic ring;

x is 1, 2 or 3.

15. The compound of item 14, wherein

16. The compound of item 15, wherein

R ⁴ Is phosphoramidate and R ¹² Is CH ₃ 、CH ₂ F、CF ₂ H or CF ₃ 。

17. The compound of item 15, wherein:

y is NR ¹ R ² ，R ¹ Is methyl and R ² Is hydrogen or methyl.

18. The compound of item 15, wherein R ²² F.

19. The compound of item 15, wherein R ²² Is OR (OR) ²⁵ 。

20. The compound of item 15, wherein:

R ²² is NR (NR) ⁵ R ⁶ And R is ⁶ Is hydrogen.

21. The compound of item 15, wherein:

R ²² is NR (NR) ⁵ R ⁶ 。

22. The compound of item 15, wherein:

R ²² selected from Cl, br, CN, N ₃ 、C ₁ -C ₆ Alkyl, C ₂ -C ₆ Alkenyl, C ₂ -C ₆ Alkynyl, - (C) ₁ -C ₂ Alkyl) (C) ₃ -C ₆ Cycloalkyl) - (C) ₀ -C ₂ Alkyl) (C) ₃ -C ₆ Heterocycle) - (C) ₀ -C ₂ Alkyl) (aryl) - (C) ₀ -C ₂ Alkyl) (heteroaryl); ONHC (=o) OR ²³ 、-NHOR ²⁴ 、-OR ²⁵ and-SR ²⁵ 。

23. The compound of item 15, wherein:

R ²² is NR (NR) ⁵ R ⁶ The method comprises the steps of carrying out a first treatment on the surface of the And

R ⁵ and R is ⁶ Is hydrogen.

24. A compound of formula III or a pharmaceutically acceptable salt thereof

Wherein:

R ⁷ is hydrogen, C _1-6 An alkyl group; c (C) _3-7 Cycloalkyl; heteroaryl, heterocycle, or aryl, wherein phenyl or naphthyl is optionally substituted with: c (C) _1-6 Alkyl, C _2-6 Alkenyl, C _2-6 Alkynyl, C _1-6 Alkoxy, F, cl, br, I, nitro, cyano, C _1-6 Haloalkyl, - (CH) ₂ ) _1-6 COOH、-(CH ₂ ) _1-6 COOC _1-6 Alkyl, -N (R) ⁷ ') ₂ 、C _1-6 Amido, NHSO ₂ C _1-6 Alkyl, -SO ₂ N(R ⁷ ') ₂ 、COR ^7" and-SO ₂ C _1-6 An alkyl group; (R) ⁷ ' independently hydrogen or C _1-6 An alkyl group; r is R ^7" is-OR ¹¹ or-N (R) ⁷ ) ₂ )；

R ^9a and R is ^9b Is (i) independently selected from hydrogen, C _1-6 Alkyl, cycloalkyl, - (CH) ₂ ) _c (NR ⁹ ') ₂ 、C _1-6 Hydroxyalkyl, -CH ₂ SH、-(CH ₂ ) ₂ S(O)(Me、-(CH ₂ ) ₃ NHC(＝NH)NH ₂ (lH-indol-3-yl) methyl, (1H-imidazol-4-yl) methyl, - (CH) ₂ ) _c COR ^9" Aryl and aryl (C) _1-3 Alkyl) -, which aryl group may be optionally substituted with a group selected from: hydroxy, C _1-6 Alkyl, C _1-6 Alkoxy, halogen, nitro and cyano; (ii) R is R ^9a And R is ^9b Are all C _1-6 An alkyl group; (iii) R is R ^9a And R is ^9b Together are (CH) ₂ ) _r Thereby forming a spiro ring; (iv) R is R ^9a Is hydrogen, and R ^9b And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising adjacent N and C atoms, (v) R ^9b Is hydrogen and R ^9a And R is ⁸ Together are (CH) ₂ ) _n Thereby forming a cyclic ring comprising contiguous N and C atoms, wherein C is 1 to 6, N is 2 to 4, R is 2 to 5, and wherein R ⁹ ' independently hydrogen or C _1-6 Alkyl and R ^9" is-OR ¹¹ or-N (R) ¹¹ ') ₂ )；(vi)R ^9a Is hydrogen and R ^9b Is hydrogen, CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH, or lower cycloalkyl; or (vii) R ^9a Is CH ₃ 、CH ₂ CH ₃ 、CH(CH ₃ ) ₂ 、CH ₂ CH(CH ₃ ) ₂ 、CH(CH ₃ )CH ₂ CH ₃ 、CH ₂ Ph、CH ₂ -indol-3-yl, -CH ₂ CH ₂ SCH ₃ 、CH ₂ CO ₂ H、CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ COOH、CH ₂ CH ₂ C(O)NH ₂ 、CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ 、-CH ₂ CH ₂ CH ₂ NHC(NH)NH ₂ 、CH ₂ Imidazol-4-yl, CH ₂ OH、CH(OH)CH ₃ 、CH ₂ ((4'-OH)-Ph)、CH ₂ SH or lower cycloalkyl and R ^9b Is hydrogen;

R ¹⁰ is hydrogen, optionally alkoxyDi (lower alkyl) -amino or halogen substituted C _1-6 Alkyl, C _1-6 Haloalkyl, C _3-7 Cycloalkyl, heterocycloalkyl, aminoacyl, aryl, for example phenyl, heteroaryl, for example pyridinyl, substituted aryl or substituted heteroaryl;

R ¹¹ is optionally substituted C _1-6 Alkyl, optionally substituted cycloalkyl; optionally substituted C _2-6 Alkynyl, optionally substituted C _2-6 Alkenyl or optionally substituted acyl groups including, but not limited to, C (O) (C _1-6 An alkyl group);

therein Y, R ³ 、R ¹² And R is ²² Defined in item 14.

25. The compound of item 24, wherein

Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is hydrogen and R ¹² Is CH ₃ 。

26. The compound of item 24, wherein

Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is methyl and R ¹² Is CH ₃ 。

27. The compound of item 24, wherein

Y is NR ¹ R ² ，R ¹ Is methyl, R ² Is cyclopropyl, R ³ Is hydrogen, R ⁷ Is phenyl, R ⁸ Is hydrogen, R ^9a Is hydrogen, R ^9b Is methyl, R ¹⁰ Is isopropyl and R ¹² Is CH ₃ 。

28. The compound of item 24, wherein

Y is NR ¹ R ² ，R ¹ Is CH ₃ ，R ¹² Is CH ₃ And R is ²² F.

29. A compound of formula IV or a pharmaceutically acceptable salt thereof

Wherein the variables Y, R ³ 、R ⁷ 、R ⁸ 、R ^9a 、R ^9b 、R ¹⁰ And R is ²² Defined in item 24.

30. A compound of formula V:

31. A compound of formula VI or a pharmaceutically acceptable salt thereof

Wherein the ratio is

Y、R ³ 、R ⁴ And R is ¹² Defined for item 24;

R ⁴¹ is F, cl, OR ³ 、N ₃ 、NH ₂ Or CN.

32. A compound of formula VII or a pharmaceutically acceptable salt thereof:

wherein the ratio is

Y、R ³ 、R ⁴ 、R ¹² 、R ²² And R is ⁴¹ Defined for items 1, 24 and 31.

33. A pharmaceutical composition comprising an effective amount of a compound of item 1 in a pharmaceutically acceptable carrier for treating HCV in a host.

34. A pharmaceutical composition comprising an effective amount of a compound of item 9 in a pharmaceutically acceptable carrier for treating HCV in a host.

35. A pharmaceutical composition comprising an effective amount of a compound of item 14 in a pharmaceutically acceptable carrier for treating HCV in a host.

36. A pharmaceutical composition comprising an effective amount of a compound of any one of items 24 or 29-32 in a pharmaceutically acceptable carrier for treating HCV in a host.

37. The pharmaceutical composition of item 33, wherein the composition is suitable for oral delivery.

38. The pharmaceutical composition of item 34, wherein the composition is suitable for oral delivery.

39. The pharmaceutical composition of item 35, wherein the composition is suitable for oral delivery.

40. The pharmaceutical composition of item 36, wherein the composition is suitable for oral delivery.

41. A method for treating a hepatitis c infection or a condition resulting from a hepatitis c infection in a host in need thereof, comprising administering an effective amount of a compound of item 1, optionally in a pharmaceutically acceptable carrier.

42. A method for treating a hepatitis c infection or a condition resulting from a hepatitis c infection in a host in need thereof, comprising administering an effective amount of a compound of item 9, optionally in a pharmaceutically acceptable carrier.

43. A method for treating a hepatitis c infection or a condition resulting from a hepatitis c infection in a host in need thereof, comprising administering an effective amount of a compound of item 14, optionally in a pharmaceutically acceptable carrier.

44. A method for treating a hepatitis c infection or a condition resulting from a hepatitis c infection in a host in need thereof, comprising administering an effective amount of a compound of any one of items 24 or 29-32, optionally in a pharmaceutically acceptable carrier.

45. The method of clause 41, wherein the compound is administered transdermally.

46. The method of item 41, wherein the compound is administered via controlled release.

47. The method of item 41, wherein the compound is administered intravenously.

48. The method of item 41, wherein the infection caused by hepatitis C is an antibody positive and antigen positive disorder, viral-based chronic hepatitis, liver cancer caused by advanced hepatitis C, cirrhosis or fatigue.

49. The method of clause 41, further comprising administering the compound in combination with another anti-HCV agent.

50. The method of clause 49, wherein the additional anti-HCV agent is selected from protease inhibitors; NS5A inhibitors; additional NS5B polymerase inhibitors; non-substrate (allosteric) inhibitors; interferon alpha-2 a, which may be pegylated; ribavirin; an helicase inhibitor; antisense oligodeoxynucleotides (S-ODNs); an aptamer; nuclease-resistant ribozymes; iRNA; antibodies to HCV; partial antibodies to HCV; and domain antibodies to HCV.

51. The method of clause 50, wherein the protease inhibitor is selected from the group consisting of telaprevir, buspiran, cimiravir, and paritaprevir.

52. The method of clauses 41-51, wherein the host is a human.

53. The compound of the formula

Wherein R is ⁴ Is a stable phosphate prodrug, or a pharmaceutically acceptable salt thereof.

54. Pharmaceutical composition comprising the following compounds

Wherein R is ⁴ Is a stable phosphate prodrug; or a pharmaceutically acceptable salt thereof.

55. The compound of the formula

56. Pharmaceutical composition comprising the following compounds

57. A compound of the structure: :

58. Pharmaceutical composition comprising the following compounds

59. A compound selected from the following structures:

in the form of an isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer, or a mixture thereof;

wherein R is ¹ 、R ² 、R ⁵ And R is ⁶ As defined in item 14.

60. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein R is ¹ And R is ² As defined in item 1.

61. A compound selected from the following structures:

wherein R is ¹ 、R ² 、R ⁵ And R is ⁶ As defined in item 14;

in the form of an isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer, or a mixture thereof.

62. Pharmaceutical compositions comprising the following compounds or pharmaceutically acceptable salts thereof

Wherein R is ¹ 、R ² 、R ⁵ 、R ⁶ 、R ⁷ And R is ¹⁰ As defined in items 9 and 14.

63. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ As defined in item 9;

64. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein R is ⁷ And R is ¹⁰ Is defined in item 9.

65. A compound selected from the following structures:

wherein R is ^3C 、R ⁷ And R is ¹⁰ As defined in items 1 and 9;

66. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

67. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

68. Pharmaceutical compositions comprising the following compounds or pharmaceutically acceptable salts thereof

Wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

69. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and wherein R ^3C Selected from alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl in the form of an isolated phosphorus R or S enantiomer or a mixture thereof, the isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer.

70. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

71. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

72. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

73. A compound selected from the following structures:

Wherein R is ^3C 、R ⁷ And R is ¹⁰ As defined in items 1 and 9;

74. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

75. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

76. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

77. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl), -O-alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl in the form of an isolated phosphorus R or S enantiomer or a mixture thereof, the isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer.

78. A pharmaceutical composition comprising the structure:

wherein R is ⁷ And R is ¹⁰ As defined in items 1 and 9; and R is ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl, or a pharmaceutically acceptable salt thereof.

79. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

80. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

81. A compound selected from the following structures:

82. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl.

83. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

84. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

85. A compound selected from the following structures:

wherein R is ^3C 、R ⁷ And R is ¹⁰ As defined in items 1 and 9;

which is in the form of the isolated phosphorus S enantiomer.

86. Pharmaceutical compositions comprising the following compounds or pharmaceutically acceptable salts thereof

Wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

87. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

in the form of the isolated phosphorus S enantiomer, or a pharmaceutically acceptable salt thereof.

88. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

89. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Selected from alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl in the form of an isolated phosphorus R or S enantiomer or a mixture thereof, the isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer.

90. A pharmaceutical composition comprising the structure:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Is or a pharmaceutically acceptable salt thereof.

91. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

92. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

93. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C In the form of an isolated phosphorus R or S enantiomer having at least 90% of the designated enantiomer, or a mixture thereof.

94. A pharmaceutical composition comprising the following compounds:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl;

or a pharmaceutically acceptable salt thereof.

95. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ ,N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

96. A pharmaceutical composition comprising the following compound or pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

97. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ Defined in items 1 and 9, and R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl

98. A pharmaceutical composition comprising the structure:

wherein R is ⁷ And R is ¹⁰ As defined in items 1 and 9; r is R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl

Or a pharmaceutically acceptable salt thereof.

99. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

100. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

101. A compound selected from the following structures:

wherein R is ⁷ And R is ¹⁰ As defined in items 1 and 9; and R is ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl

102. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

103. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

104. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

105. A compound selected from the following structures:

wherein R is ^3C 、R ⁷ And R is ¹⁰ As defined in items 1 and 9;

106. A pharmaceutical composition comprising the structure:

wherein R is ⁷ And R is ¹⁰ As defined in items 1 and 9; r is R ^3C Is alkyl, alkenyl, alkynyl, - (C) ₀ -C ₂ ) (cycloalkyl) - (C) ₀ -C ₂ ) (heterocyclyl) - (C) ₀ -C ₂ ) (aryl) - (C) ₀ -C ₂ ) (heteroaryl) and-O-alkyl or a pharmaceutically acceptable salt thereof.

107. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ As defined in item 9;

108. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

Wherein the method comprises the steps of

R ²² Selected from F and OR ²⁵ ；

R ⁷ And R is ¹⁰ Is defined in item 9.

109. A compound selected from the following structures:

wherein R is ^3C 、R ⁷ And R is ¹⁰ As defined in items 1 and 9;

110. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein R is ^3C 、R ⁷ And R is ¹⁰ Defined for items 1 and 9.

111. A compound selected from the following structures:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ As defined in item 9;

112. A pharmaceutical composition comprising the following structure or a pharmaceutically acceptable salt thereof:

wherein the method comprises the steps of

R ²² Selected from F, OR ²⁵ 、N ₃ Or CN;

R ⁷ and R is ¹⁰ Is defined in item 9.

113. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein R ³ H.

114. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein R ⁴ H.

115. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein R ⁴ Is mono-, di-or triphosphate.

116. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein the phosphorus has an S-configuration.

117. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein the phosphorus has the R-configuration.

118. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein the amino acid has a D-configuration.

119. The compound of any one of clauses 1, 9, 14, 24, or 29-32, wherein the amino acid has an L-configuration.

120. The compound of any one of clauses 14, 24, 29, 30, or 32, wherein R ²² Selected from the group consisting of chloro, bromo, fluoro, cyano, azido, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl, 1-dimethylpropyl, 2-dimethylpropyl, 3-methylbutyl, 1-ethylpropyl, vinyl, allyl, 1-butynyl, 2-butynyl, ethynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, - (CH) ₂ ) -cyclopropyl, - (CH) ₂ ) -cyclobutyl, - (CH) ₂ ) Cyclopentyl, - (CH) ₂ ) -cyclohexyl, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, pyrazolidine, piperidine, oxetane, thiane, - (CH) ₂ ) Aziridine, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Ethylene oxide, - (CH) ₂ ) Azetidine, - (CH) ₂ ) Oxetane, - (CH) ₂ ) Thietane, - (CH) ₂ ) Pyrrolidine, - (CH) ₂ ) Tetrahydrofuran, - (CH) ₂ ) Tetrahydrothiophene, - (CH) ₂ ) Pyrazolidines, - (CH) ₂ ) Piperidine, - (CH) ₂ ) -oxacyclohexane, - (CH) ₂ ) -thiacyclohexane, phenyl, pyridinyl, -ONHC (=o) OCH ₃ 、-ONHC(＝O)OCH ₂ CH ₃ 、-NHOH、NHOCH ₃ 、-OCH ₃ 、OC ₂ H ₅ 、-OPh、OCH ₂ Ph、-SCH ₃ 、-SC ₂ H ₅ 、-SPh、SCH ₂ Ph、-NH(CH ₂ ) ₂ NH ₂ 、-NH(CH ₂ ) ₂ N(CH ₃ ) ₂ 、-NHNH ₂ 、-NHNHCH ₃ 、-N＝NH、-N＝NCH ₃ 、-N＝NCH ₂ CH ₃ 、-NHC(O)NHNH ₂ 、-NHC(S)NHNH ₂ 、-C(O)NHNH ₂ 、-NHSO ₂ CH ₃ 、-NHSO ₂ CH ₂ CH ₃ 、-SO ₂ NHCH ₃ 、-SO ₂ N(CH ₃ ) ₂ 、-C(O)NH ₂ 、-C(O)NHCH ₃ 、-C(O)N(CH ₃ ) ₂ 、-CO ₂ CH ₃ 、-CO ₂ CH ₂ CH ₃ 、-CO ₂ Ph、-CO ₂ CH ₂ Ph、-SO ₂ CH ₃ 、-SO ₂ CH ₂ CH ₃ 、-SO ₂ Ph、-SO ₂ CH ₂ Ph、-P(O)H(OH)、-P(O)H(OCH ₃ )、-P(O)(OH)(OH)、-P(O)(OH)(OCH ₃ )、-P(O)(OCH ₃ )(OCH ₃ )、-P(O)(OH)(NH ₂ )、-P(O)(OH)(NHCH ₃ )、-P(O)(OH)N(CH ₃ ) ₂ 、-NHC(O)CH ₃ 、-NHC(O)CH ₂ CH ₃ 、-NHC(O)CH(CH ₃ ) ₂ 、-NHC(O)OCH ₃ 、-NHC(O)OCH ₂ CH ₃ 、-NHC(O)OCH(CH ₃ ) ₂ 、-NHC(O)OCH ₂ CH ₂ CH ₃ 、-NHC(O)OCH ₂ CH ₂ CH ₂ CH ₃ or-NHC (O) OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ 。

121. The compound of item 31, wherein R ⁴¹ Is fluorine.

122. The compound of item 31, wherein R ⁴¹ Is chlorine.

123. The compound of item 31, wherein R ⁴¹ Is hydroxyl.

124. The compound of item 32, wherein R ⁴¹ Is fluorine.

125. The compound of item 32, wherein R ⁴¹ Is chlorine.

126. The compound of item 32, wherein R ⁴¹ Is hydroxyl.

127. A compound of any one of items 1, 9, 14, 24 or 29-32, and pharmaceutically acceptable salts and prodrugs thereof, for use in the treatment or prevention of a hepatitis c virus infection. .

128. Use of a compound of any one of clauses 1, 9, 14, 24 or 29-32, and pharmaceutically acceptable salts and prodrugs thereof, in the manufacture of a medicament for treating a hepatitis c virus infection.

129. A process for the preparation of a medicament intended for therapeutic use in the treatment of hepatitis c virus infection, characterized in that a compound of any one of items 1, 9, 14, 24 or 29-32 is used in the preparation.

130. A pharmaceutical formulation comprising a therapeutically effective host amount of a compound of any one of items 1, 9, 14, 24 or 29-32, or a pharmaceutically acceptable salt or prodrug thereof, together with a pharmaceutically acceptable carrier or diluent.

131. A compound selected from:

wherein R is ⁷ 、R ¹⁰ 、R ²² And R is ⁴¹ As defined in items 9, 14 and 31.

132. The compound of the formula or the pharmaceutically acceptable salt thereof

Wherein R4 is a stable phosphate prodrug; and

R ¹ 、R ² 、R ²² and R is ⁴¹ As defined in items 1, 9, 14 and 31.

133. A compound or pharmaceutically acceptable salt thereof selected from:

wherein R4 is a stable phosphate prodrug; and

R ¹ 、R ² 、R ²² and R is ⁴¹ As defined in items 1, 9, 14 and 31.

Claims

1. The compound of the formula or the pharmaceutically acceptable salt thereof

Wherein the compound is at least 90% free of the opposite phosphorus R-enantiomer.

2. The compound according to claim 1, wherein the compound is at least 98% free of the opposite phosphorus R-enantiomer.

3. The compound according to claim 1, wherein the compound is at least 99% free of the opposite phosphorus R-enantiomer.

4. The compound of the formula or the pharmaceutically acceptable salt thereof

Wherein the compound is at least 90% free of the opposite phosphorus S-enantiomer.

5. The compound according to claim 4, wherein the compound is at least 98% free of the opposite phosphorus S-enantiomer.

6. The compound according to claim 4, wherein the compound is at least 99% free of the opposite phosphorus S-enantiomer.

7. A pharmaceutical composition comprising a compound of the formula:

8. the pharmaceutical composition according to claim 7, comprising a compound of the formula:

9. the pharmaceutical composition according to claim 8, wherein the compound is at least 90% free of the opposite phosphorus R-enantiomer.

10. The pharmaceutical composition according to claim 8, wherein the compound is at least 98% free of the opposite phosphorus R-enantiomer.

11. The pharmaceutical composition according to claim 8, wherein the compound is at least 99% free of the opposite phosphorus R-enantiomer.

12. The pharmaceutical composition according to any one of claims 7-11, which is in an oral dosage form.

13. The pharmaceutical composition according to claim 12, wherein the oral dosage form is a solid dosage form.

14. The pharmaceutical composition according to claim 13, wherein the solid dosage form is a tablet or capsule.

15. The pharmaceutical composition according to claim 12, wherein the oral dosage form is a liquid dosage form.

16. The pharmaceutical composition according to claim 15, wherein the liquid dosage form is a suspension or solution.

17. The pharmaceutical composition according to any one of claims 7-11, which is an intravenous formulation.

18. The pharmaceutical composition according to any one of claims 7-11, which is a parenteral formulation.

19. The pharmaceutical composition according to claim 7, comprising a compound of the formula:

20. the pharmaceutical composition according to claim 19, wherein the compound is at least 90% free of the opposite phosphorus S-enantiomer.

21. The pharmaceutical composition according to claim 19, wherein the compound is at least 98% free of the opposite phosphorus S-enantiomer.

22. The pharmaceutical composition according to claim 19, wherein the compound is at least 99% free of the opposite phosphorus S-enantiomer.

23. The pharmaceutical composition according to any one of claims 19-22, which is in an oral dosage form.

24. The pharmaceutical composition according to claim 23, wherein the oral dosage form is a solid dosage form.

25. The pharmaceutical composition according to claim 24, wherein the solid dosage form is a tablet or capsule.

26. The pharmaceutical composition according to claim 23, wherein the oral dosage form is a liquid dosage form.

27. The pharmaceutical composition according to claim 26, wherein the liquid dosage form is a suspension or solution.

28. The pharmaceutical composition according to any one of claims 19-22, which is an intravenous formulation.

29. The pharmaceutical composition according to any one of claims 19-22, which is a parenteral formulation.

30. A pharmaceutical composition comprising a compound of the formula:

31. a pharmaceutical composition according to claim 30 comprising a compound of the formula:

32. the pharmaceutical composition according to claim 31, wherein the compound is at least 90% free of the opposite phosphorus R-enantiomer.

33. The pharmaceutical composition according to claim 31, wherein the compound is at least 99% free of the opposite phosphorus R-enantiomer.

34. A pharmaceutical composition according to claim 30 comprising a compound of the formula:

35. the pharmaceutical composition according to claim 34, wherein the compound is at least 90% free of the opposite phosphorus S-enantiomer.

36. The pharmaceutical composition according to claim 34, wherein the compound is at least 99% free of the opposite phosphorus S-enantiomer.

37. The pharmaceutical composition according to any one of claims 30-36, which is in an oral dosage form.

38. The pharmaceutical composition according to claim 37, wherein the oral dosage form is a solid dosage form.

39. The pharmaceutical composition according to claim 38, wherein the solid dosage form is a tablet or capsule.

40. The pharmaceutical composition according to claim 37, wherein the oral dosage form is a liquid dosage form.

41. The pharmaceutical composition according to claim 40, wherein the liquid dosage form is a suspension or solution.

42. The pharmaceutical composition according to any one of claims 30-36, which is an intravenous formulation.

43. The pharmaceutical composition according to any one of claims 30-36, which is a parenteral formulation.